{"id":27017,"date":"2024-01-03T21:41:19","date_gmt":"2024-01-03T19:41:19","guid":{"rendered":"https:\/\/hho-bulgaria.com\/effects-of-hydrogen-rich-water-on-small-intestine-in-piglets\/"},"modified":"2024-01-29T21:17:31","modified_gmt":"2024-01-29T19:17:31","slug":"effects-of-hydrogen-rich-water-on-small-intestine-in-piglets","status":"publish","type":"post","link":"https:\/\/hho-bulgaria.com\/en\/effects-of-hydrogen-rich-water-on-small-intestine-in-piglets\/","title":{"rendered":"Effects of Hydrogen-rich Water on Small Intestine in Piglets"},"content":{"rendered":"<div class=\"jig-ncbiinpagenav\" data-jigconfig=\"smoothScroll: false, allHeadingLevels: ['h2'], headingExclude: ':hidden,.nomenu'\" id=\"ui-ncbiinpagenav-1\">\n<div class=\"fm-sec half_rhythm no_top_margin\">\n<div class=\"fm-flexbox\">\n<div class=\"fm-citation\">\n<div class=\"citation-default\">\n<div class=\"part1\"><span role=\"menubar\"><a href=\"#\" role=\"menuitem\" aria-expanded=\"false\" aria-haspopup=\"true\">J Anim Sci Biotechnol.<\/a><\/span> 2019; 10: 9. <\/div>\n<div class=\"part2\"><span class=\"fm-vol-iss-date\">Published online 2019 Feb 13. <\/span>  <span class=\"doi\"><span>doi:&nbsp;<\/span><a href=\"\/\/doi.org\/10.1186%2Fs40104-019-0320-2\" target=\"_blank\" rel=\"noopener noreferrer\" ref=\"reftype=other&amp;article-id=6373143&amp;issue-id=327480&amp;journal-id=1872&amp;FROM=Article%7CFront%20Matter&amp;TO=Content%20Provider%7CCrosslink%7CDOI\">10.1186\/s40104-019-0320-2<\/a><\/span><\/div>\n<\/div>\n<\/div>\n<div class=\"fm-ids\">\n<div class=\"fm-citation-pmcid\"><span class=\"fm-citation-ids-label\">PMCID: <\/span><span>PMC6373143<\/span><\/div>\n<div class=\"fm-citation-pmid\">PMID: <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/30805184\">30805184<\/a><\/div>\n<\/div>\n<\/div>\n<h1 class=\"content-title\">Morphological and molecular response of small intestine to lactulose and hydrogen-rich water in female piglets fed <em>Fusarium<\/em> mycotoxins contaminated diet<\/h1>\n<div class=\"half_rhythm\">\n<div class=\"contrib-group fm-author\"><a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Ji%20X%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139880436000496\" co-class=\"co-affbox\">Xu Ji<\/a>,<sup>1<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Zhang%20Q%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139880435433936\" co-class=\"co-affbox\">Qing Zhang<\/a>,<sup>1<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Zheng%20W%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139880445124640\" co-class=\"co-affbox\">Weijiang Zheng<\/a>,<sup><img decoding=\"async\" src=\"\/corehtml\/pmc\/pmcgifs\/corrauth.gif\" alt=\"corresponding author\"><\/sup><sup>1<\/sup> and  <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Yao%20W%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139880439370304\" co-class=\"co-affbox\">Wen Yao<\/a><sup>1,<\/sup><sup>2<\/sup><\/div>\n<div style=\"display:none\" class=\"contrib-group aff-tip\">\n<div id=\"_co_idm139880436000496\">\n<h3 class=\"no_margin\">Xu Ji<\/h3>\n<p><sup>1<\/sup>Laboratory of Gastrointestinal Microbiology, Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal Health, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095 Jiangsu China <\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Ji%20X%5BAuthor%5D\">Xu Ji<\/a><\/div>\n<\/div>\n<div id=\"_co_idm139880435433936\">\n<h3 class=\"no_margin\">Qing Zhang<\/h3>\n<p><sup>1<\/sup>Laboratory of Gastrointestinal Microbiology, Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal Health, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095 Jiangsu China <\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Zhang%20Q%5BAuthor%5D\">Qing Zhang<\/a><\/div>\n<\/div>\n<div id=\"_co_idm139880445124640\">\n<h3 class=\"no_margin\">Weijiang Zheng<\/h3>\n<p><sup>1<\/sup>Laboratory of Gastrointestinal Microbiology, Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal Health, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095 Jiangsu China <\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Zheng%20W%5BAuthor%5D\">Weijiang Zheng<\/a><\/div>\n<\/div>\n<div id=\"_co_idm139880439370304\">\n<h3 class=\"no_margin\">Wen Yao<\/h3>\n<p><sup>1<\/sup>Laboratory of Gastrointestinal Microbiology, Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal Health, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095 Jiangsu China <\/p>\n<p><sup>2<\/sup>Key Lab of Animal Physiology and Biochemistry, Ministry of Agriculture, Nanjing, 210095 Jiangsu China <\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Yao%20W%5BAuthor%5D\">Wen Yao<\/a><\/div>\n<\/div>\n<\/div>\n<\/div>\n<div class=\"half_rhythm\">\n<div class=\"togglers fm-copyright-license\"><a href=\"#\" class=\"pmctoggle\" rid=\"idm139880435326240_ai\">Author information<\/a> <a href=\"#\" class=\"pmctoggle\" rid=\"idm139880435326240_an\">Article notes<\/a> <a href=\"#\" class=\"pmctoggle\" rid=\"idm139880435326240_cpl\">Copyright and License information<\/a> <a href=\"\/pmc\/about\/disclaimer\/\" style=\"margin-left: 1em\">PMC Disclaimer<\/a><\/div>\n<div class=\"fm-authors-info hide half_rhythm\" id=\"idm139880435326240_ai\" style=\"display:none\">\n<div class=\"fm-affl\" id=\"Aff1\"><sup>1<\/sup>Laboratory of Gastrointestinal Microbiology, Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal Health, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095 Jiangsu China <\/div>\n<div class=\"fm-affl\" id=\"Aff2\"><sup>2<\/sup>Key Lab of Animal Physiology and Biochemistry, Ministry of Agriculture, Nanjing, 210095 Jiangsu China <\/div>\n<div><span class=\"fm-affl\">Xu Ji, <\/span><span class=\"fm-affl\"><span class=\"email-label\">Email: <\/span><a href=\"mailto:dev@null\" data-email=\"moc.liamg@ecnahcuxij\" class=\"oemail\">moc.liamg@ecnahcuxij<\/a><\/span>.<\/div>\n<div><a href=\"#article-aaff-info\">Contributor Information<\/a>.<\/div>\n<div><sup><img decoding=\"async\" src=\"\/corehtml\/pmc\/pmcgifs\/corrauth.gif\" alt=\"corresponding author\"><\/sup>Corresponding author.<\/div>\n<\/div>\n<div class=\"fm-article-notes hide half_rhythm\" id=\"idm139880435326240_an\" style=\"display:none\">\n<div class=\"fm-pubdate half_rhythm\">Received 2018 Aug 27; Accepted 2019 Jan 9.<\/div>\n<\/div>\n<div class=\"permissions half_rhythm hide\" id=\"idm139880435326240_cpl\" style=\"display:none\">\n<div class=\"fm-copyright half_rhythm\"><a href=\"\/pmc\/about\/copyright\/\">Copyright<\/a> \u00a9 The Author(s). 2019<\/div>\n<div class=\"license half_rhythm\"><strong>Open Access<\/strong>This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (<a href=\"http:\/\/creativecommons.org\/licenses\/by\/4.0\/\" data-ga-action=\"click_feat_suppl\" ref=\"reftype=extlink&amp;article-id=6373143&amp;issue-id=327480&amp;journal-id=1872&amp;FROM=Article%7CFront%20Matter&amp;TO=External%7CLink%7CURI\" target=\"_blank\" rel=\"noopener\">http:\/\/creativecommons.org\/licenses\/by\/4.0\/<\/a>), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (<a href=\"http:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/\" data-ga-action=\"click_feat_suppl\" ref=\"reftype=extlink&amp;article-id=6373143&amp;issue-id=327480&amp;journal-id=1872&amp;FROM=Article%7CFront%20Matter&amp;TO=External%7CLink%7CURI\" target=\"_blank\" rel=\"noopener\">http:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/<\/a>) applies to the data made available in this article, unless otherwise stated.<\/div>\n<\/div>\n<\/div>\n<div id=\"pmclinksbox\" class=\"links-box whole_rhythm hidden\" role=\"complementary\" aria-label=\"Related or updated information about this article.\"><\/div>\n<\/div>\n<div class=\"sec\"><\/div>\n<div id=\"ass-data\" class=\"tsec fm-sec whole_rhythm\" data-section=\"Featured_PMC_Datacitation\">\n<h2 class=\"nomenu\">Associated Data<\/h2>\n<dl data-count=\"2\" class=\"box-data-suppmats whole_rhythm no_bottom_margin\">\n<dt><a href=\"#\" rid=\"data-suppmats\" data-ga-action=\"click_feat_toggler\" data-ga-label=\"Supplementary Materials\" class=\"pmctoggle\">Supplementary Materials<\/a><\/dt>\n<dd id=\"data-suppmats\" style=\"display: none;\">\n<div class=\"half_rhythm\">\n<div class=\"caption half_rhythm no_bottom_margin\"><strong>Additional file 1: <\/strong><strong>Table S1.<\/strong> Ingredient composition and nutrient contents of control and experimental diets. (DOCX 20 kb)<\/div>\n<div><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/bin\/40104_2019_320_MOESM1_ESM.docx\" data-ga-action=\"click_feat_suppl\">40104_2019_320_MOESM1_ESM.docx<\/a><span style=\"color:gray\"> (21K)<\/span><\/div>\n<div class=\"small guid\">GUID:&nbsp;B8368665-088C-4BF0-B56D-816F19601D52<\/div>\n<\/div>\n<div class=\"half_rhythm\">\n<div class=\"caption half_rhythm no_bottom_margin\"><strong>Additional file 2: <\/strong><strong>Table S2.<\/strong> List of primers used in this study. (DOCX 21 kb)<\/div>\n<div><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/bin\/40104_2019_320_MOESM2_ESM.docx\" data-ga-action=\"click_feat_suppl\">40104_2019_320_MOESM2_ESM.docx<\/a><span style=\"color:gray\"> (23K)<\/span><\/div>\n<div class=\"small guid\">GUID:&nbsp;47AD14AA-D9AD-4778-BB2F-EB0CE76C6D81<\/div>\n<\/div>\n<\/dd>\n<\/dl>\n<dl data-length=\"125\" class=\"box-data-avail whole_rhythm no_bottom_margin\">\n<dt><a href=\"#\" rid=\"data-avl-stmnt\" data-ga-action=\"click_feat_toggler\" data-ga-label=\"Data Availability Statement\" class=\"pmctoggle\">Data Availability Statement<\/a><\/dt>\n<dd id=\"data-avl-stmnt\" style=\"display: none;\">\n<p class=\"p p-first-last\">The datasets used and\/or analysed during the current study are available from the corresponding author on reasonable request.<\/p>\n<\/dd>\n<\/dl>\n<\/div>\n<div id=\"Abs1\" lang=\"en\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><span role=\"menubar\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"menuitem\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/span><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"Abs1title\">Abstract<\/h2>\n<p><!--article-meta--><\/p>\n<div>\n<div id=\"sec-a.n.b.m.a\" class=\"sec sec-first\">\n<h3 id=\"sec-a.n.b.m.atitle\">Background<\/h3>\n<p id=\"Par1\" class=\"p p-first-last\">Following the intake of <em>Fusarium<\/em> mycotoxin-contaminated feed, small intestines may be exposed to high levels of toxic substances that can potentially damage intestinal functions in livestock. It is well known that <em>Fusarium<\/em> mycotoxins will lead a breakdown of the normally impeccable epithelial barrier, resulting in the development of a \u201cleaky\u201d gut. H<sub>2<\/sub> administration with different methods has been proved definitely potentials to prevent serious intestinal diseases. The goal of this study is to investigate the roles of lactulose (LAC) and hydrogen-rich water (HRW) in preventing intestinal dysfunction in piglets fed <em>Fusarium<\/em> mycotoxin-contaminated feed.<\/p>\n<\/div>\n<div id=\"sec-a.n.b.m.b\" class=\"sec\">\n<h3 id=\"sec-a.n.b.m.btitle\">Methods<\/h3>\n<p id=\"Par2\" class=\"p p-first-last\">A total of 24 female piglets were evenly assigned to 4 groups: negative control (NC) group, mycotoxin-contaminated (MC) feed&nbsp;group, MC feed with LAC treatment (MC\u2009+\u2009LAC), and MC feed with HRW treatment (MC\u2009+\u2009HRW), respectively. Piglets in the NC group were fed uncontaminated control diet, while remaining piglets were fed <em>Fusarium<\/em> mycotoxin-contaminated diet. For the NC and MC groups, 10\u2009mL\/kg body weight (BW) of hydrogen-free water (HFW) was orally administrated to piglets&nbsp;twice daily; while in the MC\u2009+\u2009LAC and MC\u2009+\u2009HRW groups, piglets were treated with the same dose of LAC solution (500\u2009mg\/kg BW) and HRW&nbsp;twice daily, respectively. On d 25, serum was collected and used for biochemical analysis. Intestinal tissues were sampled for morphological examination as well as relative genes and protein expression analysis.<\/p>\n<\/div>\n<div id=\"sec-a.n.b.m.c\" class=\"sec\">\n<h3 id=\"sec-a.n.b.m.ctitle\">Results<\/h3>\n<p id=\"Par3\" class=\"p p-first-last\">Our data showed that <em>Fusarium<\/em> mycotoxins induced higher serum diamine oxidase (DAO) activities (<em>P\u2009&lt;<\/em>&nbsp;0.05), <em>D<\/em>-lactic acid levels (<em>P\u2009&lt;<\/em>&nbsp;0.01), and endotoxin status (<em>P\u2009&lt;<\/em>&nbsp;0.01), lower villus height (<em>P\u2009&lt;<\/em>&nbsp;0.01) and ratio of villus height to crypt depth (<em>P\u2009&lt;<\/em>&nbsp;0.05) in small intestine, greater apoptosis index and higher mRNA expression related to tight junctions (<em>P\u2009&lt;<\/em>&nbsp;0.05). In addition, the distribution and down-regulation of claudin-3 (CLDN3) protein in the small intestinal was also observed. As expected, oral administrations of HRW and LAC were found to remarkably provide beneficial effects against <em>Fusarium<\/em> mycotoxin-induced apoptosis and intestinal leaking. Moreover, either HRW or LAC treatments were also revealed to prevent abnormal intestinal morphological changes, disintegrate tight junctions, and restore the expression and distribution of CLDN3 protein in the small intestinal mucosal layer in female piglets that were fed <em>Fusarium<\/em> mycotoxins contaminated diet.<\/p>\n<\/div>\n<div id=\"sec-a.n.b.m.d\" class=\"sec\">\n<h3 id=\"sec-a.n.b.m.dtitle\">Conclusions<\/h3>\n<p id=\"Par4\" class=\"p p-first-last\">Our data suggest that orally administrations of HRW and LAC result in less <em>Fusarium<\/em> mycotoxin-induced apoptosis and leak in the&nbsp;small intestine. Either HRW or LAC treatments could prevent the abnormal changes of intestinal morphology and molecular response of tight junctions as well as restore the distribution and expression of CLDN3 protein of small intestinal mucosa layer in female piglets that were fed <em>Fusarium<\/em> mycotoxins contaminated diet.<\/p>\n<\/div>\n<div id=\"sec-a.n.b.m.e\" class=\"sec sec-last\">\n<h3 id=\"sec-a.n.b.m.etitle\">Electronic supplementary material<\/h3>\n<p class=\"p p-first-last\">The online version of this article (10.1186\/s40104-019-0320-2) contains supplementary material, which is available to authorized users.<\/p>\n<\/div>\n<\/div>\n<div class=\"sec\"><strong class=\"kwd-title\">Keywords: <\/strong><span class=\"kwd-text\"><em>Fusarium<\/em> mycotoxins, Hydrogen-rich water, Lactulose, Piglets, Small intestine<\/span><\/div>\n<\/div>\n<div id=\"Sec1\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"Sec1title\">Background<\/h2>\n<p id=\"Par32\" class=\"p p-first\">Trichothecenes such as deoxynivalenol (DON) and zearalenone (ZEN) are the major <em>Fusarium<\/em> mycotoxins found in maize or feed ingredients contaminated by <em>Fusarium<\/em> fungal [<a href=\"#CR1\" rid=\"CR1\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">1<\/a>]. Gastrointestinal tract (GIT) as the first physical barrier protects the body from different kinds of contaminants, is the place where mycotoxins absorption and metabolization occur. Recently, there is increasing attention on the adverse effects of <em>Fusarium<\/em> mycotoxins on the physical structure and integrity of intestines [<a href=\"#CR2\" rid=\"CR2\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">2<\/a>, <a href=\"#CR3\" rid=\"CR3\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">3<\/a>]. <em>Fusarium<\/em> mycotoxins contaminated diet has been found to alter intestinal morphology resulting in villus atrophy and reduced villi height [<a href=\"#CR4\" rid=\"CR4\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">4<\/a>], reduced nutrition absorption [<a href=\"#CR5\" rid=\"CR5\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">5<\/a>, <a href=\"#CR6\" rid=\"CR6\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">6<\/a>], affected the expression of junctional adherent protein and tight-junction protein [<a href=\"#CR7\" rid=\"CR7\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">7<\/a>]. In addition, <em>Fusarium<\/em> mycotoxins can also induce inflammation and oxidative stress in intestinal epithelial cells [<a href=\"#CR8\" rid=\"CR8\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>], accelerating cell apoptosis, thus affecting intestinal mucosa membrane integrity [<a href=\"#CR9\" rid=\"CR9\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">9<\/a>, <a href=\"#CR10\" rid=\"CR10\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">10<\/a>]. Ultimately, those changes in intestines lead to increased intestinal permeability and decreased intestinal physical and molecular function [<a href=\"#CR3\" rid=\"CR3\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">3<\/a>]. Therefore, the implementation of practical and affordable ways to reduce or remedy the toxic effects of <em>Fusarium<\/em> mycotoxins on intestinal functions are becoming increasingly important.<\/p>\n<p id=\"Par33\">Molecular hydrogen (H<sub>2<\/sub> or hydrogen gas) has many biological effects in animals, including anti-apoptotic, anti-inflammatory, and anti-oxidant [<a href=\"#CR11\" rid=\"CR11\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">11<\/a>, <a href=\"#CR12\" rid=\"CR12\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">12<\/a>]. H<sub>2<\/sub> administration with different methods has been proved to prevent serious intestinal diseases [<a href=\"#CR13\" rid=\"CR13\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">13<\/a>\u2013<a href=\"#CR15\" rid=\"CR15\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">15<\/a>]. Among the different methods of how to ingest molecular hydrogen, hydrogen-rich water\/saline is one of the most convenient and safe way. Hydrogen-rich saline administrated via tail vein was shown to ameliorate histologic damage, as well as its ability to inhibit ischemia-reperfusion (I\/R)-induced apoptosis and to promote epithelial cell proliferation in rats [<a href=\"#CR16\" rid=\"CR16\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">16<\/a>]. Luminal injection of hydrogen-rich solution also suppressed apoptosis and intestinal tissue injury in rat intestinal IR injury model [<a href=\"#CR17\" rid=\"CR17\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">17<\/a>].<\/p>\n<p id=\"Par34\">Physiologically, a huge volume of hydrogen gas is generated daily by bacteria inside the GIT during the fermentation of residual undigested carbohydrates [<a href=\"#CR18\" rid=\"CR18\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">18<\/a>]. A part of endogenous hydrogen can be further metabolized by bacteria [<a href=\"#CR19\" rid=\"CR19\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">19<\/a>], while most of them could be diffused or absorbed into the bloodstream and transported to other host organs [<a href=\"#CR20\" rid=\"CR20\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>]. Therefore, supplementation of hydrogen-producing prebiotic could be a viable solution to provide functional hydrogen to animals. The beneficial effects of endogenous H<sub>2<\/sub> produced by fructooligosaccharides, inulin, pectin, resistant starches, turmeric, and lactulose have been well explored [<a href=\"#CR21\" rid=\"CR21\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">21<\/a>\u2013<a href=\"#CR24\" rid=\"CR24\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>].Bacterial fermentation of lactulose could dramatically increase endogenous hydrogen production, which in turn protect against intestinal damage on the models of trinitrobenzene&nbsp;sulfonic acid [<a href=\"#CR25\" rid=\"CR25\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">25<\/a>] and dextran sulfate sodium (DSS) [<a href=\"#CR24\" rid=\"CR24\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>, <a href=\"#CR26\" rid=\"CR26\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">26<\/a>].<\/p>\n<p id=\"Par35\" class=\"p p-last\">To the best of our knowledge, no study has been reported to assess the effects of HRW or LAC on <em>Fusarium<\/em> mycotoxin-induced intestinal damage in piglets. In this study, we hypothesized that both HRW and LAC can partly&nbsp;reverse damages caused by <em>Fusarium<\/em> mycotoxin-contaminated diet in female piglets, helping to maintain integrity, morphology, and barrier functions of small intestines.<\/p>\n<\/div>\n<div id=\"Sec2\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"Sec2title\">Methods<\/h2>\n<div id=\"Sec3\" class=\"sec sec-first\">\n<h3 id=\"Sec3title\">Experimental diets<\/h3>\n<p id=\"Par36\" class=\"p p-first-last\"><em>Fusarium<\/em> mycotoxins contaminated or uncontaminated corn, and the two experimental diets (NC and MC diet, respectively) were prepared as previously described [<a href=\"#CR27\" rid=\"CR27\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">27<\/a>, <a href=\"#CR28\" rid=\"CR28\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">28<\/a>]. Briefly<em>, Fusarium<\/em> mycotoxin-contaminated or uncontaminated (control) corn was used to replace 44.5% of the normal maize in the&nbsp;feed. Additional file <a href=\"#MOESM1\" rid=\"MOESM1\" class=\" media\">1<\/a>: Table S1 shows the ingredients of NC and MC diets, respectively. Previously, our data on the compositions of <em>Fusarium<\/em> mycotoxins suggested a higher mycotoxins levels in MC diet than NC diet [<a href=\"#CR27\" rid=\"CR27\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">27<\/a>, <a href=\"#CR28\" rid=\"CR28\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">28<\/a>]<em>.<\/em><\/p>\n<\/div>\n<div id=\"Sec4\" class=\"sec\">\n<h3 id=\"Sec4title\">Animals, housing, and experimental design<\/h3>\n<p id=\"Par37\" class=\"p p-first\">The experimental design was previously described [<a href=\"#CR27\" rid=\"CR27\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">27<\/a>, <a href=\"#CR28\" rid=\"CR28\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">28<\/a>]. Twenty-four female piglets (Landrace \u00d7 Large \u00d7 White) from six litters (4 piglets\/litter) were used in this study with an initial average body weight of 7.25\u2009\u00b1\u20091.02\u2009kg. Piglets from each litter were equally assigned into one of the 4 treatment groups and individually housed in pens (1.2\u2009m \u00d7 2.0\u2009m) with one feeder and one nipple drinker. The piglets had <em>ab libitum<\/em> access to feed and water. The animal trial consisted of a 6-day adaption period and a 25 d experimental period. Piglets in the NC group received uncontaminated NC diet, while the MC, MC\u2009+\u2009LAC and MC\u2009+\u2009HRW groups received <em>Fusarium<\/em> mycotoxin-contaminated (MC) diet.<\/p>\n<p id=\"Par38\" class=\"p p-last\">Oral administrations of four different treatments were also as described in our previous studies [<a href=\"#CR27\" rid=\"CR27\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">27<\/a>, <a href=\"#CR28\" rid=\"CR28\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">28<\/a>]. Piglets in each group orally received their corresponding treatment twice&nbsp;daily (10:00 and 14:00\u2009h, respectively). Hydrogen-free water (HFW)(10\u2009mL\/kg BW) was orally administrated to both NC and MC groups. The HRW containing 0.6\u20130.8\u2009mmol\/L hydrogen (Beijing Hydrovita Biotechnology Company, Beijing, China) was given to the piglets by gavage in MC\u2009+\u2009HRW group. Piglets in the MC\u2009+\u2009LAC group were administrated a dose of 500\u2009mg\/kg BW lactulose solution (Duphalac, Abbott Healthcare Products, Weesp, The Netherland) dissolved in 10\u2009mL\/kg BW volume of HFW. Due to poor health status, there was one piglet removed from each MC, MC\u2009+\u2009HRW, and MC\u2009+\u2009LAC treatments, respectively.<\/p>\n<\/div>\n<div id=\"Sec5\" class=\"sec\">\n<h3 id=\"Sec5title\">Sample collection and preparation<\/h3>\n<p id=\"Par39\" class=\"p p-first-last\">On d 24, prior to morning feeding, a&nbsp;blood sample was collected from the&nbsp;anterior vena cava of each piglet. After being placed on ice for 30\u2009min followed by centrifugation at 3,000\u00d7<em>g<\/em> for 20\u2009min at 4\u2009\u00b0C, serum was harvested and stored in a pyrogen-free glass tube at \u2212\u200980\u2009\u00b0C before analysis. By the end of the experiment, piglets were fasted overnight (12\u2009h) and euthanized by an intramuscular injection of sodium pentobarbital (40\u2009mg\/kg BW) after 30\u2009min of treatments. The duodenum (5\u2009cm from the gastric cardia), jejunum (8\u2009cm before the end of jejunal Peyer\u2019s patches) and ileum (8\u2009cm from the ileal-caecal junction) segments were collected separately and fixed in 4% paraformaldehyde for 24\u2009h before histological assays. The inner linings of duodenum, jejunum, and ileum were scraped with a smooth glass coverslip to collect mucosa samples. Samples were stored in liquid nitrogen and then at \u2212\u200980\u2009\u00b0C before RNA isolation and western blot analysis.<\/p>\n<\/div>\n<div id=\"Sec6\" class=\"sec\">\n<h3 id=\"Sec6title\">Serum chemical analysis<\/h3>\n<p id=\"Par40\" class=\"p p-first-last\">Serum <em>D<\/em>-lactic acid status and diamine oxidase (DAO) activity were measured by enzymatic spectrophotometry using a commercial kit (Jiancheng Bioengineering Institute of Nanjing, Nanjing, Jiangsu, China). Free lipopolysaccharide (LPS) in the serum was measured by a chromogenic end-point Tachypleus Amebocyte Lysate assay kit (Chinese Horseshoe Crab Reagent Manufactory, Xiamen, China) with a minimum detection limit of 0.01 endotoxin units (EU)\/mL.<\/p>\n<\/div>\n<div id=\"Sec7\" class=\"sec\">\n<h3 id=\"Sec7title\">Morphological analysis of the small intestine<\/h3>\n<p id=\"Par41\" class=\"p p-first-last\">Following fixation in 4% paraformaldehyde for 24\u2009h, the intestinal segments were embedded in paraffin and 5\u2009\u03bcm sections were sectioned with a rotary microtome. Then, the sections were stained with hematoxylin and eosin (H&amp;E) and examined under a light microscope (Olympus, Tokyo, Japan). Photomicrographs were taken with an Olympus CKX31 microscope (Olympus, Tokyo, Japan). The morphometric analysis was performed on 10 randomly-selected, well-oriented villi and crypts per piglet. A computerized microscope-based image analyzer (Olympus dotslide Virtual Slide System, Tokyo, Japan) was used to determine the height of villus (from the tip of the villus to the villus-crypt junction) and crypt depth (from the crypt-villus junction to the base of the crypt).<\/p>\n<\/div>\n<div id=\"Sec8\" class=\"sec\">\n<h3 id=\"Sec8title\">Determination of apoptosis by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)<\/h3>\n<p id=\"Par42\" class=\"p p-first-last\">DNA fragments on paraffin-embedded sections of three small intestine parts (duodenum, jejunum, and ileum) were detected by One Step TUNEL Apoptosis Assay Kit (Beyotime, Nantong, China). Ten randomly selected photos were taken with a laser-scanning confocal microscope (Zeiss LSM 700 META; Jena, Germany). The total numbers of cells undergoing apoptosis were counted. The apoptosis index (AI) was calculated by the equation: AI\u2009=\u2009number of apoptotic cells \/ total number of cells \u00d7\u2009100%.<\/p>\n<\/div>\n<div id=\"Sec9\" class=\"sec\">\n<h3 id=\"Sec9title\">Quantitative gene expression analysis<\/h3>\n<p id=\"Par43\" class=\"p p-first-last\">Total RNA from the duodenum, jejunum, and ileum mucosa were isolated using FastRNA\u00ae Pro Green Kit (MP Biomedicals, USA). The yield and purity of mRNA were measured spectrophotometrically (Nanodrop 2000, Thermo Fisher, USA). Reverse transcription was conducted using a Prime Script\u2122 RT reagent Kit with gDNA Eraser (Perfect Real Time) (Takara, Dalian, China). qRT-PCR was performed using SYBR Premix Ex Taq\u2122 (Takara, Dalian, China) with the QuantStudio 5 Real-Time PCR System (Thermo Fisher, USA). Specific gene&nbsp;primers for B-cell CLL\/lymphoma 2 (<em>Bcl-2<\/em>), caspase-3 and fas cell surface death receptor (<em>FAS<\/em>), zonula occludens 1 (<em>ZO-1<\/em>), occludin (<em>OCLN<\/em>), claudin-1 (<em>CLDN1<\/em>), and claudin-3 (<em>CLDN3<\/em>) (Additional file <a href=\"#MOESM2\" rid=\"MOESM2\" class=\" media\">2<\/a>: Table S2) were detected. The real-time PCR reactions were performed using the following cycle program: precycling at 95\u2009\u00b0C for 30\u2009s followed by 40\u2009cycles of denaturization for 5\u2009s at 95\u2009\u00b0C and annealing for 30\u2009s at 60\u2009\u00b0C. \u03b2-actin was used as a&nbsp;reference gene for normalization. The relative mRNA expression levels of the target gene in comparison with the reference gene were calculated using the 2<sup>\u2013\u0394\u0394Ct<\/sup> method.<\/p>\n<\/div>\n<div id=\"Sec10\" class=\"sec\">\n<h3 id=\"Sec10title\">Western blot analysis<\/h3>\n<p id=\"Par44\" class=\"p p-first-last\">Intestinal mucosa samples were lysed using RIPA buffer (Roche, Shanghai, China). The concentrations of protein in samples were measured by the bicinchoninic acid (BCA) protein assay kit (Beyotime, Nantong, China). The total protein samples were separated through a 10% SDS polyacrylamide gel and then transferred to a nitrocellulose membrane (Boster, Wuhan, China). The&nbsp;membrane was incubated in 1:10,000 monoclonal mouse anti-beta actin (Bioworld, USA), and 1:1,000 rabbit polyclonal CLDN3 antibodies (Abcam, Shanghai, China) at 4\u2009\u00b0C overnight. Then membrane was incubated in 1:10,000 diluted horseradish peroxidase (HRP)-conjugated anti-rabbit antibody (Bioworld, USA) or 1:10,000 diluted HRP-conjugated anti-mouse antibody (Bioworld, USA) for 1\u2009h at room temperature. Tanon\u2122 High-sig ECL Western Blotting Substrate (Tanon, Shanghai, China) was applied to the membrane for 5\u2009min after secondary antibody incubation. The chemiluminescent signals were visualized by the Versa Doc\u2122 imaging system. Signal intensity was quantified using Quantity One software (Bio-Rad, USA). Protein expression levels were normalized with \u03b2-actin expression level.<\/p>\n<\/div>\n<div id=\"Sec11\" class=\"sec\">\n<h3 id=\"Sec11title\">Immunohistochemistry<\/h3>\n<p id=\"Par45\" class=\"p p-first-last\">Small intestine parts (duodenum, jejunum, and ileum) were immersed in 4% paraformaldehyde for 24\u2009h, dehydrated in ethanol, and embedded in paraffin. The tissues were sectioned into 5\u2009\u03bcm thickness on a rotary microtome. After antigen retrieval in 10\u2009mmol\/L citrate buffer (pH\u20097.5) for 3\u2009min in a microwave, paraffin sections were deparaffinized. Tissue endogenous peroxidase activity was blocked with 30% H<sub>2<\/sub>O<sub>2<\/sub> (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China) in methanol (1\u2009h). Sections were incubated with 3% bovine serum albumin (BSA) (DSBIO, Beijing, China) before overnight incubation with CLDN3 (1:200 diluted) antibody (Abcam, Shanghai, China) at 4\u2009\u00b0C. Tissue sections were then incubated with biotinylated goat&nbsp;anti-rabbit secondary antibody (1:100, Boster, Wuhan, China) followed by strept avidin-biotin complex (SABC) (1:100, Boster, Wuhan, China). Diaminobenzidine (DAB) (DSBIO, Beijing, China) solution was used to stain the prepared slides for 5\u2009min. After immunoreaction, the images were captured on each slide at Olympus CKX31 microscope (Olympus, Tokyo, Japan). The average density of positive cells was quantified using the Image-Pro Plus software (Media Cybernetics, Bethesda, MD, USA).<\/p>\n<\/div>\n<div id=\"Sec12\" class=\"sec sec-last\">\n<h3 id=\"Sec12title\">Statistical analysis<\/h3>\n<p id=\"Par46\" class=\"p p-first-last\">Statistical analysis was performed by the one-way ANOVA procedure of SPSS 18.0 (SPSS, Inc., Chicago, IL, USA, 2009). Differences among treatments were compared using Turkey-Kramer test and considered statistically significant at <em>P<\/em>\u2009&lt;\u20090.05.<\/p>\n<\/div>\n<\/div>\n<div id=\"Sec13\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"Sec13title\">Results<\/h2>\n<div id=\"Sec14\" class=\"sec sec-first\">\n<h3 id=\"Sec14title\">Status intestinal mucosal permeability<\/h3>\n<p id=\"Par47\" class=\"p p-first-last\">Compared with the NC diet, piglets fed <em>Fusarium<\/em> mycotoxins-contaminated diet (MC) had higher DAO activity (<em>P<\/em>\u2009&lt;\u20090.05), and greater concentrations of <em>D<\/em>-lactic acid and endotoxin (<em>P<\/em>\u2009&lt;\u20090.01) (Table&nbsp;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/table\/Tab1\/\" target=\"table\" class=\"fig-table-link figpopup\" rid-figpopup=\"Tab1\" rid-ob=\"ob-Tab1\" co-legend-rid=\"\" rel=\"noopener\"><span>1<\/span><\/a>). Oral administration of either HRW or LAC lower the DAO activities, <em>D<\/em>-lactic acid levels, and LPS concentrations in MC\u2009+\u2009LAC and MC\u2009+\u2009HRW piglets compared with the MC group (<em>P<\/em>\u2009&lt;\u20090.05). No difference was found among the NC, MC\u2009+\u2009LAC and MC\u2009+\u2009HRW groups in above-mentioned tests.<\/p>\n<p><!--table ft1--><!--table-wrap mode=\"anchored\" t5--><\/p>\n<div class=\"table-wrap anchored whole_rhythm\" id=\"Tab1\">\n<h3>Table 1<\/h3>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of lactulose and hydrogen-rich water on serum <em>D<\/em>-lactic acid levels, diamine oxidase (DAO) activities and endotoxin concentrations in female piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet<sup>1, 2<\/sup><\/p>\n<\/div>\n<div class=\"xtable\">\n<table frame=\"hsides\" rules=\"groups\" class=\"rendered small default_table\">\n<thead>\n<tr>\n<th rowspan=\"1\" colspan=\"1\">Item<\/th>\n<th rowspan=\"1\" colspan=\"1\">NC<\/th>\n<th rowspan=\"1\" colspan=\"1\">MC<\/th>\n<th rowspan=\"1\" colspan=\"1\">MC\u2009+\u2009LAC<\/th>\n<th rowspan=\"1\" colspan=\"1\">MC\u2009+\u2009HRW<\/th>\n<th rowspan=\"1\" colspan=\"1\">SEM<\/th>\n<th rowspan=\"1\" colspan=\"1\"><em>P<\/em>-value<\/th>\n<\/tr>\n<\/thead>\n<tbody>\n<tr>\n<td rowspan=\"1\" colspan=\"1\"><em>D<\/em>-Lactic acid, mmol\/L<\/td>\n<td rowspan=\"1\" colspan=\"1\">12.53<sup>Bb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">21.47<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">14.15<sup>Bb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">14.14<sup>Bb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.04<\/td>\n<td rowspan=\"1\" colspan=\"1\">&lt;\u20090.001<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">DAO, U\/mL<\/td>\n<td rowspan=\"1\" colspan=\"1\">13.37<sup>b<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">19.97<sup>a<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">13.52<sup>b<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">13.68<sup>b<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">0.96<\/td>\n<td rowspan=\"1\" colspan=\"1\">0.020<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">Endotoxin, EU\/L<\/td>\n<td rowspan=\"1\" colspan=\"1\">0.68<sup>Bb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.26<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">0.74<sup>Bb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">0.84<sup>ABb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">0.06<\/td>\n<td rowspan=\"1\" colspan=\"1\">0.001<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139880439204528\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/table\/Tab1\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"tblwrap-foot\">\n<p>Data are expressed as mean with standard error of mean (SEM)<\/p>\n<p><sup>(A,B,a,b)<\/sup>Means in the same row not sharing the same lower (<em>P<\/em>\u2009&lt;\u20090.05) or upper (<em>P<\/em>\u2009&lt;\u20090.01) case superscript letters are significantly different<\/p>\n<p><sup>1<\/sup> NC (negative control), basal diet; MC, <em>Fusarium<\/em> mycotoxin-contaminated diet; MC\u2009+\u2009LAC, MC diet + lactulose treatment; and MC\u2009+\u2009HRW, MC diet + hydrogen-rich water treatment<\/p>\n<p><sup>2<\/sup><br \/>\n<em>n<\/em>\u2009=\u20095<\/p>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"Sec15\" class=\"sec\">\n<h3 id=\"Sec15title\">Small intestinal morphological changes<\/h3>\n<p id=\"Par48\" class=\"p p-first\">No difference was found on crypt depth in duodenum, jejunum, and ileum among the four groups (Table&nbsp;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/table\/Tab2\/\" target=\"table\" class=\"fig-table-link figpopup\" rid-figpopup=\"Tab2\" rid-ob=\"ob-Tab2\" co-legend-rid=\"\" rel=\"noopener\"><span>2<\/span><\/a>). Compared with the NC group, piglets fed MC diet had a&nbsp;lower height of villus in duodenum, jejunum, and ileum (<em>P<\/em>\u2009&lt;\u20090.01). Compared with the MC group, both HRW and LAC treatments attenuated the reduction of villus height in small intestines (duodenum, jejunum, and ileum) caused by <em>Fusarium<\/em> mycotoxin-contaminated diet (<em>P<\/em>\u2009&lt;\u20090.01). In the duodenum and jejunum, the ratio of villus height to crypt depth in the MC group was lower than the&nbsp;NC group (<em>P<\/em>\u2009&lt;\u20090.01). Compared with the MC group, oral administrations of HRW and LAC increased the value of villus height to crypt depth (<em>P<\/em>\u2009&lt;\u20090.01) in both duodenum and jejunum. And no difference was observed among MC, MC\u2009+\u2009HRW, and MC\u2009+\u2009LAC groups for the ratio of villus height to crypt depth in&nbsp;the ileum.<\/p>\n<p><!--table ft1--><!--table-wrap mode=\"anchored\" t5--><\/p>\n<div class=\"table-wrap anchored whole_rhythm\" id=\"Tab2\">\n<h3>Table 2<\/h3>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of lactulose and hydrogen-rich water on small intestinal morphology in female piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet<sup>1, 2<\/sup><\/p>\n<\/div>\n<div class=\"xtable\">\n<table frame=\"hsides\" rules=\"groups\" class=\"rendered small default_table\">\n<thead>\n<tr>\n<th rowspan=\"1\" colspan=\"1\">Item<\/th>\n<th rowspan=\"1\" colspan=\"1\">NC<\/th>\n<th rowspan=\"1\" colspan=\"1\">MC<\/th>\n<th rowspan=\"1\" colspan=\"1\">MC\u2009+\u2009LAC<\/th>\n<th rowspan=\"1\" colspan=\"1\">MC\u2009+\u2009HRW<\/th>\n<th rowspan=\"1\" colspan=\"1\">SEM<\/th>\n<th rowspan=\"1\" colspan=\"1\"><em>P<\/em>-value<\/th>\n<\/tr>\n<\/thead>\n<tbody>\n<tr>\n<td colspan=\"7\" rowspan=\"1\">Duodenum<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">\u2003Villus height, \u03bcm<\/td>\n<td rowspan=\"1\" colspan=\"1\">386.08<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">313.51<sup>Bb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">366.65<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">360.49<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">6.86<\/td>\n<td rowspan=\"1\" colspan=\"1\">&lt;\u20090.001<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">\u2003Crypt depth, \u03bcm<\/td>\n<td rowspan=\"1\" colspan=\"1\">268.13<\/td>\n<td rowspan=\"1\" colspan=\"1\">281.75<\/td>\n<td rowspan=\"1\" colspan=\"1\">272.35<\/td>\n<td rowspan=\"1\" colspan=\"1\">272.50<\/td>\n<td rowspan=\"1\" colspan=\"1\">3.85<\/td>\n<td rowspan=\"1\" colspan=\"1\">0.678<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">\u2003Villus height: crypt depth ratio<\/td>\n<td rowspan=\"1\" colspan=\"1\">1.45<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.11<sup>Bb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.36<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.32<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">0.03<\/td>\n<td rowspan=\"1\" colspan=\"1\">0.001<\/td>\n<\/tr>\n<tr>\n<td colspan=\"7\" rowspan=\"1\">Jejunum<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">\u2003Villus height, \u03bcm<\/td>\n<td rowspan=\"1\" colspan=\"1\">390.60<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">322.41<sup>Bb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">383.75<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">380.46<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">7.03<\/td>\n<td rowspan=\"1\" colspan=\"1\">&lt;\u20090.001<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">\u2003Crypt depth, \u03bcm<\/td>\n<td rowspan=\"1\" colspan=\"1\">246.11<\/td>\n<td rowspan=\"1\" colspan=\"1\">240.94<\/td>\n<td rowspan=\"1\" colspan=\"1\">247.31<\/td>\n<td rowspan=\"1\" colspan=\"1\">231.89<\/td>\n<td rowspan=\"1\" colspan=\"1\">2.73<\/td>\n<td rowspan=\"1\" colspan=\"1\">0.172<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">\u2003Villus height: crypt depth ratio<\/td>\n<td rowspan=\"1\" colspan=\"1\">1.59<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.34<sup>Bb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.55<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.64<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">0.03<\/td>\n<td rowspan=\"1\" colspan=\"1\">&lt;\u20090.001<\/td>\n<\/tr>\n<tr>\n<td colspan=\"7\" rowspan=\"1\">Ileum<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">\u2003Villus height, \u03bcm<\/td>\n<td rowspan=\"1\" colspan=\"1\">376.88<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">334.76<sup>Bb<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">366.28<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">363.09<sup>Aa<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">4.41<\/td>\n<td rowspan=\"1\" colspan=\"1\">0.001<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">\u2003Crypt depth, \u03bcm<\/td>\n<td rowspan=\"1\" colspan=\"1\">240.35<\/td>\n<td rowspan=\"1\" colspan=\"1\">235.78<\/td>\n<td rowspan=\"1\" colspan=\"1\">240.00<\/td>\n<td rowspan=\"1\" colspan=\"1\">257.36<\/td>\n<td rowspan=\"1\" colspan=\"1\">3.36<\/td>\n<td rowspan=\"1\" colspan=\"1\">0.097<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"1\" colspan=\"1\">\u2003Villus height: crypt depth ratio<\/td>\n<td rowspan=\"1\" colspan=\"1\">1.57<sup>a<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.43<sup>ab<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.53<sup>ab<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">1.41<sup>b<\/sup><\/td>\n<td rowspan=\"1\" colspan=\"1\">0.02<\/td>\n<td rowspan=\"1\" colspan=\"1\">0.030<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139880440035984\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/table\/Tab2\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"tblwrap-foot\">\n<p>Data are expressed as mean with standard error of mean (SEM)<\/p>\n<p><sup>(A,B,a,b)<\/sup>Means in the same row not sharing the same lower (<em>P<\/em>\u2009&lt;\u20090.05) or upper (<em>P<\/em>\u2009&lt;\u20090.01) case superscript letters are significantly different<\/p>\n<p><sup>1<\/sup> NC (negative control), basal diet; MC, <em>Fusarium<\/em> mycotoxin-contaminated diet; MC\u2009+\u2009LAC, MC diet + lactulose treatment; and MC\u2009+\u2009HRW, MC diet + hydrogen-rich water treatment<\/p>\n<p><sup>2<\/sup><br \/>\n<em>n<\/em>\u2009=\u20095<\/p>\n<\/div>\n<\/div>\n<p id=\"Par49\" class=\"p p-last\">Representative morphologic observations of the intestinal tissue in the&nbsp;duodenum (Fig.&nbsp;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig1\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig1\" rid-ob=\"ob-Fig1\" co-legend-rid=\"lgnd_Fig1\" rel=\"noopener\"><span>1<\/span><\/a>a, b, c, and d), jejunum (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig1\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig1\" rid-ob=\"ob-Fig1\" co-legend-rid=\"lgnd_Fig1\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.1e,<\/span><\/span><span>1<\/span><\/a>e, f, g, and h), and ileum (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig1\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig1\" rid-ob=\"ob-Fig1\" co-legend-rid=\"lgnd_Fig1\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.1i,<\/span><\/span><span>1<\/span><\/a>i, j, k, and l) from NC, MC, MC\u2009+\u2009LAC, and MC\u2009+\u2009HRW groups are shown in Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig1\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig1\" rid-ob=\"ob-Fig1\" co-legend-rid=\"lgnd_Fig1\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2em;\">Fig.1.<\/span><\/span><span>1<\/span><\/a>. Morphology examination revealed that obvious denuded to the villi&nbsp;and&nbsp;damages were found in piglets from MC group than NC group. However,&nbsp;these morphological changes in duodenum and ileum were not seen in HRW or LAC groups.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"Fig1\" co-legend-rid=\"lgnd_Fig1\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig1\/\" target=\"figure\" rid-figpopup=\"Fig1\" rid-ob=\"ob-Fig1\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139880435237872\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig1\/\" target=\"figure\" rid-figpopup=\"Fig1\" rid-ob=\"ob-Fig1\" rel=\"noopener\"><\/a><a class=\"inline_block ts_canvas\" href=\"\/core\/lw\/2.0\/html\/tileshop_pmc\/tileshop_pmc_inline.html?title=Click%20on%20image%20to%20zoom&amp;p=PMC3&amp;id=6373143_40104_2019_320_Fig1_HTML.jpg\" target=\"tileshopwindow\" rel=\"noopener\"><\/p>\n<div class=\"ts_bar small\" title=\"Click on image to zoom\"><\/div>\n<p><img decoding=\"async\" loading=\"lazy\" alt=\"An external file that holds a picture, illustration, etc.\nObject name is 40104_2019_320_Fig1_HTML.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/40104_2019_320_Fig1_HTML.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139880435237872\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig1\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_Fig1\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig1\/\" target=\"figure\" rid-figpopup=\"Fig1\" rid-ob=\"ob-Fig1\" rel=\"noopener\">Fig. 1<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of lactulose and hydrogen-rich water on histological changes of the&nbsp;small intestine in female piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet. Representative haematoxylin &amp; eosin (H&amp;E) staining images were obtained at 200\u00d7 magnification with black bar\u2009=\u2009100\u2009\u03bcm. <strong>a<\/strong>&#8211;<strong>d<\/strong> duodenum tissue images. <strong>e<\/strong>&#8211;<strong>h<\/strong> jejunum tissue images. <strong>i<\/strong>&#8211;<strong>l<\/strong> ileum tissue images. NC (negative control), basal diet; MC, <em>Fusarium<\/em> mycotoxin-contaminated diet; MC\u2009+\u2009LAC, MC diet + lactulose treatment; and MC\u2009+\u2009HRW, MC diet + hydrogen-rich water treatment<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"Sec16\" class=\"sec\">\n<h3 id=\"Sec16title\">Small intestinal mRNA changes related to apoptosis genes<\/h3>\n<p id=\"Par50\" class=\"p p-first-last\">No difference was found on mRNA expression levels of <em>Bcl-2<\/em>, caspase-3 and <em>FAS<\/em> in the duodenum among the four groups (Fig.&nbsp;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig2\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig2\" rid-ob=\"ob-Fig2\" co-legend-rid=\"lgnd_Fig2\" rel=\"noopener\"><span>2<\/span><\/a>a). In jejunum (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig2\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig2\" rid-ob=\"ob-Fig2\" co-legend-rid=\"lgnd_Fig2\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.2b),<\/span><\/span><span>2<\/span><\/a>b), mRNA expression levels of <em>Bcl-2<\/em> and caspase-3 in the&nbsp;MC group were higher (<em>P<\/em>\u2009&lt;\u20090.05) and those in the NC group. However, mRNA expression levels of <em>Bcl-2<\/em> and caspase-3 in the MC\u2009+\u2009HRW and MC\u2009+\u2009LAC groups were lower (<em>P<\/em>\u2009&lt;\u20090.01) than those in the MC group. <em>FAS<\/em> mRNA expression levels in jejunum had no difference among the four groups. In ileum (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig2\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig2\" rid-ob=\"ob-Fig2\" co-legend-rid=\"lgnd_Fig2\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.2),<\/span><\/span><span>2<\/span><\/a>), mRNA expression levels of <em>Bcl-2<\/em> and <em>FAS<\/em> were not different among the four treatment groups. MC group showed higher (<em>P<\/em>\u2009&lt;\u20090.05) caspase-3 gene mRNA expression level than NC group. However, ileum caspase-3 mRNA expression levels were not different among the NC, MC\u2009+\u2009HRW and MC\u2009+\u2009LAC groups.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"Fig2\" co-legend-rid=\"lgnd_Fig2\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig2\/\" target=\"figure\" rid-figpopup=\"Fig2\" rid-ob=\"ob-Fig2\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139880440761200\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig2\/\" target=\"figure\" rid-figpopup=\"Fig2\" rid-ob=\"ob-Fig2\" rel=\"noopener\"><\/a><a class=\"inline_block ts_canvas\" href=\"\/core\/lw\/2.0\/html\/tileshop_pmc\/tileshop_pmc_inline.html?title=Click%20on%20image%20to%20zoom&amp;p=PMC3&amp;id=6373143_40104_2019_320_Fig2_HTML.jpg\" target=\"tileshopwindow\" rel=\"noopener\"><\/p>\n<div class=\"ts_bar small\" title=\"Click on image to zoom\"><\/div>\n<p><img decoding=\"async\" loading=\"lazy\" alt=\"An external file that holds a picture, illustration, etc.\nObject name is 40104_2019_320_Fig2_HTML.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/40104_2019_320_Fig2_HTML.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139880440761200\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig2\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_Fig2\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig2\/\" target=\"figure\" rid-figpopup=\"Fig2\" rid-ob=\"ob-Fig2\" rel=\"noopener\">Fig. 2<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of lactulose and hydrogen-rich water on relative mRNA gene expression levels related to apoptosis in&nbsp;the duodenum (<strong>a<\/strong>), jejunum (<strong>b<\/strong>), and ileum (<strong>c<\/strong>) of female&nbsp;piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet. Each column represents the mean values (<em>n<\/em>\u2009=\u20095), with their standard deviation (SD) represented by vertical bars. Letters above the bars not sharing the same lower (<em>P<\/em>\u2009&lt;\u20090.05) or upper (<em>P<\/em>\u2009&lt;\u20090.01) case superscript are significantly different. <em>Bcl-2<\/em>\u2009=\u2009B-cell CLL\/lymphoma 2, <em>FAS<\/em>\u2009=\u2009Fas cell surface death receptor. NC\u2009=\u2009basal diet; MC\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet; MC\u2009+\u2009LAC\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet with lactulose treatment. MC\u2009+\u2009HRW\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet with hydrogen-rich water treatment<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"Sec17\" class=\"sec\">\n<h3 id=\"Sec17title\">Intestinal apoptosis status detected by TUNEL<\/h3>\n<p id=\"Par51\" class=\"p p-first\">Representative observations of apoptosis in the duodenum (Fig.&nbsp;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig3\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig3\" rid-ob=\"ob-Fig3\" co-legend-rid=\"lgnd_Fig3\" rel=\"noopener\"><span>3<\/span><\/a> a, a2, a3, and a4), jejunum (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig3\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig3\" rid-ob=\"ob-Fig3\" co-legend-rid=\"lgnd_Fig3\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.3<\/span><\/span><span>3<\/span><\/a> a5, a6, a7, and a8), and ileum (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig3\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig3\" rid-ob=\"ob-Fig3\" co-legend-rid=\"lgnd_Fig3\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.3a9,<\/span><\/span><span>3<\/span><\/a>a9, a10, a11, and a12) from NC, MC, MC\u2009+\u2009LAC, and MC\u2009+\u2009HRW groups were shown in Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig3\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig3\" rid-ob=\"ob-Fig3\" co-legend-rid=\"lgnd_Fig3\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2em;\">Fig.3a.<\/span><\/span><span>3<\/span><\/a>a. Following in situ labeling of duodenum, jejunum, and ileum mucosal epithelium, stained epithelial cells from the jejunum and ileum were undergoing apoptosis in piglets fed MC diet were seen under microscopic examination. A number of TUNEL-positive cells were also found in piglets from MC\u2009+\u2009HRW and MC\u2009+\u2009LAC groups, respectively.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"Fig3\" co-legend-rid=\"lgnd_Fig3\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig3\/\" target=\"figure\" rid-figpopup=\"Fig3\" rid-ob=\"ob-Fig3\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139880441856112\"><img decoding=\"async\" loading=\"lazy\" class=\"fig-image\" alt=\"An external file that holds a picture, illustration, etc.\nObject name is 40104_2019_320_Fig3_HTML.jpg\" title=\"An external file that holds a picture, illustration, etc.\nObject name is 40104_2019_320_Fig3_HTML.jpg\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/40104_2019_320_Fig3_HTML.jpg\"><\/div>\n<p><\/a><\/p>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139880441856112\" style=\"display: none;\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig3\/\" target=\"figure\" rid-figpopup=\"Fig3\" rid-ob=\"ob-Fig3\" rel=\"noopener\"><\/a><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig3\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_Fig3\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig3\/\" target=\"figure\" rid-figpopup=\"Fig3\" rid-ob=\"ob-Fig3\" rel=\"noopener\">Fig. 3<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of lactulose and hydrogen-rich water on the percentage of apoptosis intestinal epithelial cells by the TUNEL assay in female&nbsp;piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet. <strong>a<\/strong>&nbsp;Representative TUNEL stained paraffin sections from the&nbsp;duodenum (a1\u20134), jejunum (a5\u20138), and ileum (a9\u201312) tissue (original magnification, 400\u00d7). <strong>b<\/strong> Effects of lactulose and hydrogen-rich water on the percentage of small intestine apoptosis ratio in piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet. Each column represents the mean values (<em>n<\/em>\u2009=\u20095), with their standard deviation (SD) represented by vertical bars. Letters above the bars not sharing the same lower (<em>P<\/em>\u2009&lt;\u20090.05) or upper (<em>P<\/em>\u2009&lt;\u20090.01) case superscript letters are significantly different. NC\u2009=\u2009basal diet; MC\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet; MC\u2009+\u2009HRW\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet with hydrogen-rich water treatment; MC\u2009+\u2009LAC\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet with lactulose treatment<\/p>\n<\/div>\n<\/div>\n<\/div>\n<p id=\"Par52\" class=\"p p-last\">The apoptosis index for the quantification of TUNEL-positive cells is shown in Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig3\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig3\" rid-ob=\"ob-Fig3\" co-legend-rid=\"lgnd_Fig3\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2em;\">Fig.3b.<\/span><\/span><span>3<\/span><\/a>b. In&nbsp;the duodenum, apoptotic index among the four groups was not different. In jejunum, an increase of apoptosis index was found in piglets fed MC diet than those that fed NC diet (<em>P<\/em>\u2009&lt;\u20090.01). Compared with the MC group, only LAC treatment lowered the jejunum apoptosis index (<em>P<\/em>\u2009&lt;\u20090.05). In ileum, MC group had a higher apoptosis index than any of NC, MC\u2009+\u2009LAC, and MC\u2009+\u2009HRW groups (<em>P<\/em>\u2009&lt;\u20090.01). No difference in AI was seen among NC, MC\u2009+\u2009LAC and MC\u2009+\u2009HRW groups.<\/p>\n<\/div>\n<div id=\"Sec18\" class=\"sec\">\n<h3 id=\"Sec18title\">mRNA changes of genes related to small intestinal barrier function<\/h3>\n<p id=\"Par53\" class=\"p p-first\">In duodenum (Fig.&nbsp;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig4\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig4\" rid-ob=\"ob-Fig4\" co-legend-rid=\"lgnd_Fig4\" rel=\"noopener\"><span>4<\/span><\/a>a), mRNA expression levels of <em>ZO-1<\/em>, <em>OCLN,<\/em> and <em>CLDN1<\/em> had no difference among the four treatment groups. mRNA expression levels of <em>CLDN3<\/em> in the MC group was higher than NC, MC\u2009+\u2009LAC, and MC\u2009+\u2009HRW groups (<em>P<\/em>\u2009&lt;\u20090.01). No difference was detected among the NC, MC\u2009+\u2009LAC, and MC\u2009+\u2009HRW groups.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"Fig4\" co-legend-rid=\"lgnd_Fig4\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig4\/\" target=\"figure\" rid-figpopup=\"Fig4\" rid-ob=\"ob-Fig4\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139880439409392\"><img decoding=\"async\" loading=\"lazy\" class=\"fig-image\" alt=\"An external file that holds a picture, illustration, etc.\nObject name is 40104_2019_320_Fig4_HTML.jpg\" title=\"An external file that holds a picture, illustration, etc.\nObject name is 40104_2019_320_Fig4_HTML.jpg\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/40104_2019_320_Fig4_HTML.jpg\"><\/div>\n<p><\/a><\/p>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139880439409392\" style=\"display: none;\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig4\/\" target=\"figure\" rid-figpopup=\"Fig4\" rid-ob=\"ob-Fig4\" rel=\"noopener\"><\/a><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig4\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_Fig4\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig4\/\" target=\"figure\" rid-figpopup=\"Fig4\" rid-ob=\"ob-Fig4\" rel=\"noopener\">Fig. 4<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of lactulose and hydrogen-rich water on relative mRNA gene expression levels related to tight junctions in&nbsp;the duodenum (<strong>a<\/strong>), jejunum (<strong>b<\/strong>), and ileum (<strong>c<\/strong>) of female&nbsp;piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet. Each column represents the mean values (<em>n<\/em>\u2009=\u20095), with their standard deviation (SD) represented by vertical bars. Letters above the bars not sharing the same lower (<em>P<\/em>\u2009&lt;\u20090.05) or upper (<em>P<\/em>\u2009&lt;\u20090.01) case superscript letters are significantly different. NC\u2009=\u2009basal diet; MC\u2009=\u2009mycotoxin-contaminated diet; MC\u2009+\u2009LAC\u2009=\u2009mycotoxin-contaminated diet with lactulose treatment. MC\u2009+\u2009HRW\u2009=\u2009mycotoxin-contaminated diet with hydrogen-rich water treatment<\/p>\n<\/div>\n<\/div>\n<\/div>\n<p id=\"Par54\">In jejunum (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig4\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig4\" rid-ob=\"ob-Fig4\" co-legend-rid=\"lgnd_Fig4\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.4b),<\/span><\/span><span>4<\/span><\/a>b), no difference was found in mRNA expression levels of <em>ZO-1<\/em> and <em>CLDN1<\/em> among the four treatment groups. MC diet stimulated the increase of <em>OCLN<\/em> and <em>CLDN3<\/em> mRNA expression levels compared with NC diet (<em>P<\/em>\u2009&lt;\u20090.01). Both MC\u2009+\u2009LAC and MC\u2009+\u2009HRW groups had decreased <em>OCLN<\/em> and <em>CLDN3<\/em> mRNA expression levels compared with the&nbsp;MC group (<em>P<\/em>\u2009&lt;\u20090.05).<\/p>\n<p id=\"Par55\" class=\"p p-last\">In ileum (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig4\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig4\" rid-ob=\"ob-Fig4\" co-legend-rid=\"lgnd_Fig4\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.4c),<\/span><\/span><span>4<\/span><\/a>c), mRNA expression level of <em>ZO-1<\/em> in the&nbsp;MC group was higher than its expression level in NC, MC\u2009+\u2009HRW and MC\u2009+\u2009LAC groups (<em>P<\/em>\u2009&lt;\u20090.01). No difference in <em>ZO-1<\/em> mRNA expression was seen among the MC, MC\u2009+\u2009LAC and MC\u2009+\u2009HRW groups. mRNA expression level of <em>CLDN3<\/em> in the&nbsp;MC group was higher than it in the NC, MC\u2009+\u2009LAC, and MC\u2009+\u2009HRW groups (<em>P<\/em>\u2009&lt;\u20090.01). And <em>CLDN3<\/em> mRNA expression level in MC\u2009+\u2009LAC group was higher than the&nbsp;NC group (<em>P<\/em>\u2009&lt;\u20090.05). However, mRNA expression levels of <em>OCLN<\/em> and <em>CLDN1<\/em> had no difference among the four treatment groups.<\/p>\n<\/div>\n<div id=\"Sec19\" class=\"sec\">\n<h3 id=\"Sec19title\">Relative expression of CLDN3 protein in the&nbsp;small intestine<\/h3>\n<p id=\"Par56\" class=\"p p-first-last\">Using western blotting technique, no difference was found on the expression levels of CLDN3 protein in duodenum among the four treatment groups&nbsp;(Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig5\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig5\" rid-ob=\"ob-Fig5\" co-legend-rid=\"lgnd_Fig5\" rel=\"noopener\"><span>5<\/span><\/a>). In both jejunum and ileum&nbsp;(Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig5\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig5\" rid-ob=\"ob-Fig5\" co-legend-rid=\"lgnd_Fig5\" rel=\"noopener\"><span>5<\/span><\/a>), CLDN3 protein levels were lower in the&nbsp;MC group than in NC, MC\u2009+\u2009LAC, and MC\u2009+\u2009HRW groups (<em>P<\/em>\u2009&lt;\u20090.05). No difference of CLDN3 protein expression level was detected among NC, MC\u2009+\u2009LAC, and MC\u2009+\u2009HRW groups in&nbsp;the jejunum&nbsp;(Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig5\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig5\" rid-ob=\"ob-Fig5\" co-legend-rid=\"lgnd_Fig5\" rel=\"noopener\"><span>5<\/span><\/a>). While MC\u2009+\u2009LAC and MC\u2009+\u2009HRW groups had a higher ileum CLDN3 protein expression levels (<em>P<\/em>\u2009&lt;\u20090.05) than MC group&nbsp;(Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig5\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig5\" rid-ob=\"ob-Fig5\" co-legend-rid=\"lgnd_Fig5\" rel=\"noopener\"><span>5<\/span><\/a>).<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"Fig5\" co-legend-rid=\"lgnd_Fig5\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig5\/\" target=\"figure\" rid-figpopup=\"Fig5\" rid-ob=\"ob-Fig5\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139880439137408\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig5\/\" target=\"figure\" rid-figpopup=\"Fig5\" rid-ob=\"ob-Fig5\" rel=\"noopener\"><\/a><a class=\"inline_block ts_canvas\" href=\"\/core\/lw\/2.0\/html\/tileshop_pmc\/tileshop_pmc_inline.html?title=Click%20on%20image%20to%20zoom&amp;p=PMC3&amp;id=6373143_40104_2019_320_Fig5_HTML.jpg\" target=\"tileshopwindow\" rel=\"noopener\"><\/p>\n<div class=\"ts_bar small\" title=\"Click on image to zoom\"><\/div>\n<p><img decoding=\"async\" loading=\"lazy\" alt=\"An external file that holds a picture, illustration, etc.\nObject name is 40104_2019_320_Fig5_HTML.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/40104_2019_320_Fig5_HTML.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139880439137408\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig5\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_Fig5\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig5\/\" target=\"figure\" rid-figpopup=\"Fig5\" rid-ob=\"ob-Fig5\" rel=\"noopener\">Fig. 5<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of lactulose and hydrogen-rich water on claudin-3(CLDN3) protein expression in duodenum, jejunum, and ileum of female&nbsp;piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet. Each column represents the mean values (<em>n<\/em>\u2009=\u20095), with their standard deviation (SD) represented by vertical bars. Letters above the bars not sharing the same lower (<em>P<\/em>\u2009&lt;\u20090.05) or upper (<em>P<\/em>\u2009&lt;\u20090.01) case superscript letters are significantly different. NC\u2009=\u2009basal diet; MC\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet; MC\u2009+\u2009HRW\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet with hydrogen-rich water treatment; MC\u2009+\u2009LAC\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet with lactulose treatment<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"Sec20\" class=\"sec sec-last\">\n<h3 id=\"Sec20title\">Distribution of CLDN3 protein in the&nbsp;surface of small intestinal<\/h3>\n<p id=\"Par57\" class=\"p p-first\">The localization of CLDN3 protein status of the duodenum, jejunum, and ileum in four treatment groups was determined via immunohistochemistry assay. Strong positive immunoreactivity results were obtained in duodenum&nbsp;(Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" co-legend-rid=\"lgnd_Fig6\" rel=\"noopener\"><span>6<\/span><\/a> a1, a2, a3, and a4), jejunum&nbsp;(Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" co-legend-rid=\"lgnd_Fig6\" rel=\"noopener\"><span>6<\/span><\/a> a5, a6, a7, and a8), and ileum&nbsp;(Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" co-legend-rid=\"lgnd_Fig6\" rel=\"noopener\"><span>6<\/span><\/a> a9, a10, a11, and a12) indicated by brown staining (Fig.&nbsp;<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" co-legend-rid=\"lgnd_Fig6\" rel=\"noopener\"><span>6<\/span><\/a>). CLDN3 protein can be seen on the full surface of villi especially clustered at the tips of villi (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" co-legend-rid=\"lgnd_Fig6\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.6a).<\/span><\/span><span>6a<\/span><\/a>). All three small intestinal villi in&nbsp;the NC group exhibited a continuous pattern of CLDN3 protein lining, which indicated good intestinal integrity (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" co-legend-rid=\"lgnd_Fig6\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.6<\/span><\/span><span>6<\/span><\/a> a1, a5, and a9, respectively). <em>Fusarium<\/em> mycotoxin-contaminated diet was associated with a&nbsp;disturbed and irregular distribution of CLDN3 proteins in duodenum, jejunum, and ileum samples (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" co-legend-rid=\"lgnd_Fig6\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.6<\/span><\/span><span>6<\/span><\/a> a2, a6, and a10), respectively. In the meanwhile, MC\u2009+\u2009HRW (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" co-legend-rid=\"lgnd_Fig6\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.6<\/span><\/span><span>6<\/span><\/a> a4, a8, and a12) and MC\u2009+\u2009LAC (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" co-legend-rid=\"lgnd_Fig6\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.6<\/span><\/span><span>6<\/span><\/a> a3, a7, and a11) groups had less irregular CLDN3 distribution than MC group.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"Fig6\" co-legend-rid=\"lgnd_Fig6\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139880483180464\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" rel=\"noopener\"><\/a><a class=\"inline_block ts_canvas\" href=\"\/core\/lw\/2.0\/html\/tileshop_pmc\/tileshop_pmc_inline.html?title=Click%20on%20image%20to%20zoom&amp;p=PMC3&amp;id=6373143_40104_2019_320_Fig6_HTML.jpg\" target=\"tileshopwindow\" rel=\"noopener\"><\/p>\n<div class=\"ts_bar small\" title=\"Click on image to zoom\"><\/div>\n<p><img decoding=\"async\" loading=\"lazy\" alt=\"An external file that holds a picture, illustration, etc.\nObject name is 40104_2019_320_Fig6_HTML.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/40104_2019_320_Fig6_HTML.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139880483180464\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_Fig6\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" rel=\"noopener\">Fig. 6<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of lactulose and hydrogen-rich water on claudin-3(CLDN3) protein density levels of the&nbsp;small intestine in female&nbsp;piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet.&nbsp;<strong>a<\/strong> Representative immunohistochemical staining images of CLDN3 protein in the&nbsp;duodenum (a1\u20134), jejunum (a5\u20138), and ileum (a9\u201312) were obtained at 400\u00d7 magnification with red bar\u2009=\u200950\u2009\u03bcm. <strong>b<\/strong> Mean density (arbitrary units) in the&nbsp;small intestine of piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet. Each column represents the mean values (<em>n<\/em>\u2009=\u20095), with their standard deviation (SD) represented by vertical bars. Letters above the bars not sharing the same lower (<em>P<\/em>\u2009&lt;\u20090.05) or upper (<em>P<\/em>\u2009&lt;\u20090.01) case superscript letters are significantly different. NC\u2009=\u2009basal diet; MC\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet; MC\u2009+\u2009HRW\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet with hydrogen-rich water treatment; MC\u2009+\u2009LAC\u2009=\u2009<em>Fusarium<\/em> mycotoxin-contaminated diet with lactulose treatment<\/p>\n<\/div>\n<\/div>\n<\/div>\n<p id=\"Par58\" class=\"p p-last\">The abundance of CLDN3 protein in the duodenum, jejunum, and ileum was also measured (Fig. <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/figure\/Fig6\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"Fig6\" rid-ob=\"ob-Fig6\" co-legend-rid=\"lgnd_Fig6\" rel=\"noopener\"><span style=\"position: relative;text-decoration:none;\">\u200b<span class=\"figpopup-sensitive-area\" style=\"left: -2.5em;\">(Fig.6b).<\/span><\/span><span>6<\/span><\/a>b). Compared with NC group, MC group had a&nbsp;lower density of CLDN3 protein in duodenum, jejunum, and ileum (<em>P<\/em>\u2009&lt;\u20090.05). In the&nbsp;duodenum, oral administration of either HRW or LAC did not alter CLDN3 protein expression density compared with the MC group. However, oral administrations of either HRW or LAC were found increased the CLDN3 expression levels when compared with the MC group in both jejunum and ileum (<em>P<\/em>\u2009&lt;\u20090.05).<\/p>\n<\/div>\n<\/div>\n<div id=\"Sec21\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"Sec21title\">Discussion<\/h2>\n<p id=\"Par59\" class=\"p p-first\">GIT is not only responsible for food ingestion, digestion, energy and nutrients absorption, but also an essential barrier preventing the passage of harmful intraluminal substances from the external environment [<a href=\"#CR29\" rid=\"CR29\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">29<\/a>]. Following the intake of <em>Fusarium<\/em> mycotoxin-contaminated feed, GIT can be exposed to high levels of toxic substances that consequentially damage intestinal functions [<a href=\"#CR3\" rid=\"CR3\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">3<\/a>]. HRW is known could ameliorate intestinal histologic damage and injury [<a href=\"#CR14\" rid=\"CR14\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">14<\/a>, <a href=\"#CR15\" rid=\"CR15\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">15<\/a>]. LAC has also been shown protective effects against different models of intestinal damage through endogenous hydrogen [<a href=\"#CR22\" rid=\"CR22\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>\u2013<a href=\"#CR24\" rid=\"CR24\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>]. Therefore, we hypothesized that oral administrations of either HRW or LAC could partially reverse the damages in small intestinal tracts caused by <em>Fusarium<\/em> mycotoxins in piglets.<\/p>\n<p id=\"Par60\">The absorption of mycotoxins and their fate in the intestinal tract suggest that the gut epithelium is repeatedly exposed to these toxins, and at higher levels than other tissues [<a href=\"#CR2\" rid=\"CR2\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">2<\/a>\u2013<a href=\"#CR4\" rid=\"CR4\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">4<\/a>]. Serum <em>D<\/em>-lactic acid levels, DAO activities, and endotoxin status are useful markers for measuring the permeability of the intestinal mucosa, intestinal injury and reperfusion insults [<a href=\"#CR30\" rid=\"CR30\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">30<\/a>]. In this study, <em>Fusarium<\/em> mycotoxin-contaminated feed induced abnormal intestinal tissue structural changes and altered intestinal permeability. A previous report [<a href=\"#CR31\" rid=\"CR31\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">31<\/a>] indicated that DON exposure not only caused a reduction in transepithelial electrical resistance (TEER) of intestinal epithelial cell monolayers but also increased the permeability of epithelial intestinal cell monolayers to bacteria. In vivo experiment also shown that serum concentrations of <em>D<\/em>-lactic acid and DAO were also elevated in piglets challenged with 4\u2009mg\/kg deoxynivalenol [<a href=\"#CR32\" rid=\"CR32\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">32<\/a>]. Those findings are consistent with our results that piglets fed <em>Fusarium<\/em> mycotoxin-contaminated feed had displayed higher serum <em>D<\/em>-lactic acid levels, DAO activities and LPS concentrations than those in the NC group.<\/p>\n<p id=\"Par61\">It was reported that 2% hydrogen inhalation could attenuate I\/R injury induced histopathological mucosal erosion and increased gut permeability via its antioxidant effects in rats [<a href=\"#CR33\" rid=\"CR33\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">33<\/a>]. Intra-peritoneal injection of hydrogen-rich saline (10\u2009mL\/kg) was also found to maintain the body weight, attenuate the severity of necrotizing enterocolitis (NEC), and prevent the&nbsp;increase of serum DAO in a neonatal rat model of NEC [<a href=\"#CR34\" rid=\"CR34\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">34<\/a>]. In this study, compared with the&nbsp;MC group, lower levels of serum <em>D<\/em>-lactic acid, DAO activities, and endotoxin concentrations were detected in both MC\u2009+\u2009LAC and MC\u2009+\u2009HRW groups. In our previous studies with exactly same piglets, oral administrations of HRW or LAC showed higher hydrogen concentrations in plasma and intestine [<a href=\"#CR27\" rid=\"CR27\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">27<\/a>, <a href=\"#CR28\" rid=\"CR28\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">28<\/a>]. Although the underlying mechanisms of the HRW and LAC exert their protective effects on the gut permeability remains unknown, the&nbsp;antioxidative property of molecular hydrogen might shed the light on further discovering the underlying mechanism. Therefore, molecular hydrogen may be a good and novel candidate agent to reduce the side effects caused by <em>Fusarium<\/em> mycotoxins in piglets.<\/p>\n<p id=\"Par62\">Changes of intestinal structure, including villus height, crypt depth and the ratio of villus height to crypt depth ratio are considered sensitive indicators of&nbsp;the intestine that reacts to the presence of harmful substances in feed [<a href=\"#CR35\" rid=\"CR35\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">35<\/a>]. Chronic (5-week&nbsp;long) ingestion of a diet contaminated with DON (3\u2009mg\/kg) alone or together with fumonisins (6\u2009mg\/kg) induced morphological changes in pig intestine, these changes included atrophy and fusion of villi, decreased of villi height, and cell proliferation in the jejunum [<a href=\"#CR4\" rid=\"CR4\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">4<\/a>]. In addition, feeding 5-week-old piglets for 28 d with multiple <em>Fusarium<\/em> toxin-contaminated feed caused decreased of villus height and crypt depth in both jejunum and ileum [<a href=\"#CR36\" rid=\"CR36\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">36<\/a>]. In our study, piglets fed <em>Fusarium<\/em> mycotoxin-contaminated diet had shorter villus height and a&nbsp;lower ratio of villus height to crypt depth in duodenum, jejunum, and ileum, suggesting that the physiological architecture of small intestine was compromised. These results might be partially accounted for higher serum DAO activities, <em>D<\/em>-lactic acids and LPS levels in the&nbsp;MC group than NC group due to increase intestinal permeability caused by small intestinal structure damage [<a href=\"#CR31\" rid=\"CR31\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">31<\/a>].<\/p>\n<p id=\"Par63\">Furthermore, histological examination demonstrated that both HRW and LAC prevented <em>Fusarium<\/em> mycotoxin-induced mucosal structural changes in duodenum, jejunum, and ileum. A previous study reported that jugular venous cannula infusion of hydrogen-rich saline (5\u2009mL\/kg) significantly reduced the mucosa injury caused by IR, preventing shortened villi, loss of villous epithelium and prominent mucosa neutrophil infiltration in the small intestine of Sprague-Dawley rats [<a href=\"#CR15\" rid=\"CR15\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">15<\/a>]. Lactulose was able to reduce the colonic damage [<a href=\"#CR24\" rid=\"CR24\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>, <a href=\"#CR26\" rid=\"CR26\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">26<\/a>] of DSS and trinitrobenzenesulfonic acid [<a href=\"#CR25\" rid=\"CR25\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">25<\/a>] models by increasing hydrogen production. So, it is not surprising to see that both HRW and LAC oral administrations have&nbsp;shown similar beneficial effects against <em>Fusarium<\/em> mycotoxin-induced intestinal damage in piglets. Since the intestinal is the key organ to digest feed and absorb nutrients, the protective effects of HRW and LAC on small intestinal morphology can be beneficial to improve growth performance in piglets [<a href=\"#CR27\" rid=\"CR27\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">27<\/a>].<\/p>\n<p id=\"Par64\">Several studies indicated that mycotoxins belong to trichothecenes can cause apoptosis in bone marrow, marcophages, Peyer\u2019s patches and thymus [<a href=\"#CR37\" rid=\"CR37\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">37<\/a>, <a href=\"#CR38\" rid=\"CR38\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">38<\/a>]. Aflatoxin B1 (0.3\u2009mg\/kg) could induce the increase of apoptotic thymocyte by up-regulation mRNA expression level of <em>Bax<\/em> and caspase and down-regulation mRNA expression level of <em>Bcl-2<\/em> [<a href=\"#CR39\" rid=\"CR39\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">39<\/a>]. However, the effects of <em>Fusarium<\/em> mycotoxins on the apoptosis of small intestine were rarely explored in weaning piglets. In our study, feeding <em>Fusarium<\/em> mycotoxin-contaminated diet up-regulated <em>Bcl-2<\/em> and caspase-3 mRNA expression in jejunum, and caspase-3 expression in ileum. <em>Fusarium<\/em> mycotoxin-induced apoptosis is detected by TUNEL assay with higher epithelium apoptosis ratio in jejunum and ileum in MC group.<\/p>\n<p id=\"Par65\">Molecular hydrogen has the ability to inhibit I\/R-induced oxidative stress and apoptosis and promote epithelial cell proliferation [<a href=\"#CR14\" rid=\"CR14\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">14<\/a>, <a href=\"#CR17\" rid=\"CR17\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">17<\/a>]. Hydrogen-rich saline could promote acinar cell proliferation, inhibit apoptosis and NF-\u03baB activation from <em>L<\/em>-arginine-induced acute pancreatitis in rats [<a href=\"#CR40\" rid=\"CR40\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">40<\/a>]. In our study, 10\u2009mL\/kg BW of HRW (twice&nbsp;daily) and 500\u2009mg\/kg BW of LAC (twice&nbsp;daily) significantly down-regulated the apoptosis-related gene expression in the jejunum (<em>Bcl-2<\/em> and caspase-3) and ileum (caspase-3) compared with the MC group. The protective effects of HRW and LAC against <em>Fusarium<\/em> mycotoxins were also confirmed by TUNEL assay. Sun H et al., [<a href=\"#CR41\" rid=\"CR41\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">41<\/a>] has also reported a similar finding that the activation of caspase-3 decreased remarkably in the presence of hydrogen-rich saline.<\/p>\n<p id=\"Par66\">It was also shown in our study that <em>CLDN3<\/em> mRNA expression was up-regulated by <em>Fusarium<\/em> mycotoxin-contaminated diet in the&nbsp;small intestine. Jejunum <em>OCLN<\/em> and ileum <em>ZO-1<\/em> mRNA expression levels were also up-regulated. Immunohistochemistry analysis results in small intestine sections also supported these results. These changes are also reported in other in vivo and in vitro studies. Up-regulation in mRNA expression levels of <em>CLDN3<\/em> and claudin-4 was observed in DON-exposed Caco-2 cells [<a href=\"#CR42\" rid=\"CR42\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">42<\/a>]. Low-dose (0.9\u2009mg\/kg feed), short-term exposure (10 d) of DON to piglets significantly changed the mRNA expression of different tight junction proteins in different parts of the small intestine [<a href=\"#CR43\" rid=\"CR43\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">43<\/a>]. However, no clear explanation for the contradicting results on mRNA,&nbsp;and protein expression levels of CLDN3. It can be speculated that this could be related to many factors such as exposure time, the age of piglets, and the compositions of <em>Fusarium<\/em> mycotoxins and the individual&nbsp;mycotoxin levels. Due to the replication limit, further experiments involved&nbsp;a large number of piglets and pure mycotoxins are definitely needed to be explored.<\/p>\n<p id=\"Par67\" class=\"p p-last\">The association of excessive oxidative stress and <em>Fusarium<\/em> mycotoxin-induced intestinal barrier dysfunction has also been reported [<a href=\"#CR10\" rid=\"CR10\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">10<\/a>]. Our finding suggested that oral administrations of HRW and LAC not only attenuated the morphology damage of intestine but also protected the reduction of tight junctions in the small intestines caused by <em>Fusarium<\/em> mycotoxins. Previous studies in our lab demonstrated that endogenous hydrogen gas levels in the intestines and plasma were significantly improved by HRW and LAC [<a href=\"#CR27\" rid=\"CR27\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">27<\/a>, <a href=\"#CR28\" rid=\"CR28\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">28<\/a>]. These endogenous gas may work against the side effects caused by <em>Fusarium<\/em> mycotoxins on tight junctions of the&nbsp;small intestine through its antioxidant and anti-inflammatory effects. H<sub>2<\/sub> administration with different methods can contribute to prevention of severe intestinal diseases such as transplantation [<a href=\"#CR33\" rid=\"CR33\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">33<\/a>], ischemia\/reperfusion injury [<a href=\"#CR13\" rid=\"CR13\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">13<\/a>, <a href=\"#CR17\" rid=\"CR17\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">17<\/a>, <a href=\"#CR44\" rid=\"CR44\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">44<\/a>] and colon inflammation [<a href=\"#CR13\" rid=\"CR13\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">13<\/a>, <a href=\"#CR14\" rid=\"CR14\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">14<\/a>]. In addition, hydrogen-producing prebiotic (oligosaccharides and lactulose) has been demonstrated effective in intestinal inflammation models [<a href=\"#CR24\" rid=\"CR24\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>\u2013<a href=\"#CR26\" rid=\"CR26\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">26<\/a>].&nbsp;Furthermore, intestinal microbe also plays important roles in&nbsp;regulating the development and health of small intestine [<a href=\"#CR28\" rid=\"CR28\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">28<\/a>, <a href=\"#CR45\" rid=\"CR45\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">45<\/a>]. Whether intestinal microbe plays a role in the beneficial effects of HRW and LAC in the current study remains unclear.<\/p>\n<\/div>\n<div id=\"Sec22\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"Sec22title\">Conclusions<\/h2>\n<p id=\"Par68\" class=\"p p-first-last\">In conclusion, this study demonstrated that oral administrations of HRW and LAC provided beneficial effects in reducing apoptosis of epithelium cells in small intestine, maintaining intestinal barrier, preventing intestinal morphological changes, and tight junctions disintegration, and restore the protein expression and distribution of CLDN3 in the small intestinal in female piglets fed <em>Fusarium<\/em> toxins contaminated diet. These findings provide a possible explanation for the curative effects of molecular hydrogen on <em>Fusarium<\/em> mycotoxins-induced growth depression, and a novel solution to alleviate the intestinal toxicity caused by <em>Fusarium<\/em> mycotoxins in swine production.<\/p>\n<\/div>\n<div id=\"sec-a.o.f\" class=\"tsec sec\"><a id=\"supplementary-material-sec\"><\/a><\/p>\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"sec-a.o.ftitle\">Additional files<\/h2>\n<p><!--\/article\/body\/sec\/--><\/p>\n<div id=\"Sec23\" class=\"sec sec-first-last\">\n<p class=\"p p-first-last\">\n<div class=\"sec suppmat\" id=\"MOESM1\">\n<div class=\"sup-box half_rhythm\" id=\"media-a.o.f.b.a.a.a\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/bin\/40104_2019_320_MOESM1_ESM.docx\" data-ga-action=\"click_feat_suppl\">Additional file 1:<\/a><sup>(21K, docx)<\/sup><\/p>\n<p><strong>Table S1.<\/strong> Ingredient composition and nutrient contents of control and experimental diets. (DOCX 20 kb)<\/p>\n<\/div>\n<\/div>\n<div class=\"sec suppmat\" id=\"MOESM2\">\n<div class=\"sup-box half_rhythm\" id=\"media-a.o.f.b.a.b.a\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC6373143\/bin\/40104_2019_320_MOESM2_ESM.docx\" data-ga-action=\"click_feat_suppl\">Additional file 2:<\/a><sup>(23K, docx)<\/sup><\/p>\n<p><strong>Table S2.<\/strong> List of primers used in this study. (DOCX 21 kb)<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"ack-a.p.b\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"ack-a.p.btitle\">Acknowledgements<\/h2>\n<div class=\"sec\">\n<p>The authors would like to thank Prof. Mingguo Zhou and Dr. Yabing Duan for proving <em>Fusarium graminearum<\/em> strain 2021 and preparing the conidia.<\/p>\n<div id=\"FPar1\" class=\"sec\">\n<h3 id=\"FPar1title\">Funding<\/h3>\n<p id=\"Par69\" class=\"p p-first-last\">This work was supported by grants from the National Nature Science Foundation of China (31501986), Fundamental Research Funds for Central Universities (KJQN201611 and KYDS201809) and Earmarked Fund for Jiangsu Agricultural Industry Technology System (SXGC[2018]287).<\/p>\n<\/div>\n<div id=\"FPar2\" class=\"sec sec-last\">\n<h3 id=\"FPar2title\">Availability of data and materials<\/h3>\n<p id=\"Par70\" class=\"p p-first-last\">The datasets used and\/or analysed during the current study are available from the corresponding author on reasonable request.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"glossary-a.p.a\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"glossary-a.p.atitle\">Abbreviations<\/h2>\n<div class=\"bk-sec\">\n<div>\n<table class=\"default-table glossary\">\n<tbody>\n<tr>\n<td>AI<\/td>\n<td>Apoptosis index<\/td>\n<\/tr>\n<tr>\n<td>BCA<\/td>\n<td>Bicinchoninic acid<\/td>\n<\/tr>\n<tr>\n<td>Bcl-2<\/td>\n<td>B-cell CLL\/lymphoma 2<\/td>\n<\/tr>\n<tr>\n<td>BSA<\/td>\n<td>Bovine serum albumin<\/td>\n<\/tr>\n<tr>\n<td>BW<\/td>\n<td>Body weight<\/td>\n<\/tr>\n<tr>\n<td>CLDN1<\/td>\n<td>Claudin-1<\/td>\n<\/tr>\n<tr>\n<td>CLDN3<\/td>\n<td>Claudin-3<\/td>\n<\/tr>\n<tr>\n<td>DAO<\/td>\n<td>Diamine oxidase<\/td>\n<\/tr>\n<tr>\n<td>DON<\/td>\n<td>Deoxynivalenol<\/td>\n<\/tr>\n<tr>\n<td>DSS<\/td>\n<td>Dextran sulfate sodium<\/td>\n<\/tr>\n<tr>\n<td>EU<\/td>\n<td>Endotoxin units<\/td>\n<\/tr>\n<tr>\n<td>FAS<\/td>\n<td>Fas cell surface death receptor<\/td>\n<\/tr>\n<tr>\n<td>GIT<\/td>\n<td>Gastrointestinal tract<\/td>\n<\/tr>\n<tr>\n<td>H&amp;E<\/td>\n<td>Hematoxylin and eosin<\/td>\n<\/tr>\n<tr>\n<td>HFW<\/td>\n<td>Hydrogen-free water<\/td>\n<\/tr>\n<tr>\n<td>HRW<\/td>\n<td>Hydrogen-rich water<\/td>\n<\/tr>\n<tr>\n<td>I\/R<\/td>\n<td>Ischemia-reperfusion<\/td>\n<\/tr>\n<tr>\n<td>LAC<\/td>\n<td>Lactulose<\/td>\n<\/tr>\n<tr>\n<td>LPS<\/td>\n<td>Lipopolysaccharide<\/td>\n<\/tr>\n<tr>\n<td>MC<\/td>\n<td>Mycotoxin-contaminated<\/td>\n<\/tr>\n<tr>\n<td>NC<\/td>\n<td>Negative control<\/td>\n<\/tr>\n<tr>\n<td>OCLN<\/td>\n<td>Occludin<\/td>\n<\/tr>\n<tr>\n<td>PBS<\/td>\n<td>Phosphate-buffered saline<\/td>\n<\/tr>\n<tr>\n<td>SABC<\/td>\n<td>Strept avidin-biotin complex<\/td>\n<\/tr>\n<tr>\n<td>TUNEL<\/td>\n<td>Terminal deoxynucleotidyl transferase dUTP nick end labeling<\/td>\n<\/tr>\n<tr>\n<td>ZEN<\/td>\n<td>Zearalenone<\/td>\n<\/tr>\n<tr>\n<td>ZO-1<\/td>\n<td>Zonula occludens 1<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"notes-a.p.c\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"notes-a.p.ctitle\">Authors\u2019 contributions<\/h2>\n<p>WJZ and WY designed the study. XJ and QZ conducted the experiment. XJ and QZ performed and collected the data. XJ analyzed the data. WJZ wrote the manuscript. All authors read and approved the final manuscript.<\/p>\n<\/div>\n<div id=\"notes-a.p.d\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"notes-a.p.dtitle\">Notes<\/h2>\n<div id=\"FPar3\" class=\"sec sec-first\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 id=\"FPar3title\" class=\"ui-helper-clearfix\">Ethics approval and consent to participate<\/h2>\n<p class=\"p p-first-last\">This protocol of this study was approved by the Committee of Animal Research Institute (Certification No. SYXK (Su) 2011\u20130036), Nanjing Agricultural University, China. The studies were conducted in the Animal Research Facility at Nanjing Agricultural University, China.<\/p>\n<\/div>\n<div id=\"FPar4\" class=\"sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 id=\"FPar4title\" class=\"ui-helper-clearfix\">Consent for publication<\/h2>\n<p class=\"p p-first-last\">Not applicable.<\/p>\n<\/div>\n<div id=\"FPar5\" class=\"sec sec-last\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 id=\"FPar5title\" class=\"ui-helper-clearfix\">Competing interests<\/h2>\n<p class=\"p p-first-last\">The authors declare that they have no competing interests.<\/p>\n<\/div>\n<\/div>\n<div id=\"article-aaff-info\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"article-aaff-infotitle\">Contributor Information<\/h2>\n<p><span class=\"fm-affl\">Xu Ji, <\/span><span class=\"fm-affl\"><span class=\"email-label\">Email: <\/span><a href=\"mailto:dev@null\" data-email=\"moc.liamg@ecnahcuxij\" class=\"oemail\">moc.liamg@ecnahcuxij<\/a><\/span>.<\/p>\n<p><span class=\"fm-affl\">Qing Zhang, <\/span><span class=\"fm-affl\"><span class=\"email-label\">Email: <\/span><a href=\"mailto:dev@null\" data-email=\"moc.361@eezgniqgnahz\" class=\"oemail\">moc.361@eezgniqgnahz<\/a><\/span>.<\/p>\n<p><span class=\"fm-affl\">Weijiang Zheng, <\/span><span class=\"fm-affl\">Phone: +86-25-8439-9830, <span class=\"email-label\">Email: <\/span><a href=\"mailto:dev@null\" data-email=\"nc.ude.uajn@gnaijiewgnehz\" class=\"oemail\">nc.ude.uajn@gnaijiewgnehz<\/a><\/span>.<\/p>\n<p><span class=\"fm-affl\">Wen Yao, <\/span><span class=\"fm-affl\"><span class=\"email-label\">Email: <\/span><a href=\"mailto:dev@null\" data-email=\"nc.ude.uajn@pj76newoay\" class=\"oemail\">nc.ude.uajn@pj76newoay<\/a><\/span>.<\/p>\n<\/div>\n<div id=\"Bib1\" class=\"tsec sec\">\n<div class=\"goto jig-ncbiinpagenav-goto-container\"><a class=\"tgt_dark page-toc-label jig-ncbiinpagenav-goto-heading\" href=\"#\" title=\"Go to other sections in this page\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Go to:<\/a><\/div>\n<h2 class=\"head no_bottom_margin ui-helper-clearfix\" id=\"Bib1title\">References<\/h2>\n<div class=\"ref-list-sec sec\" id=\"reference-list\">\n<div class=\"ref-cit-blk half_rhythm\" id=\"CR1\">1. <span class=\"element-citation\">Shi H, Li S, Bai Y, Prates LL, Lei Y, Yu P. 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href=\"https:\/\/scholar.google.com\/scholar_lookup?journal=J+Toxicol+Environ+Health+B+Crit+Rev&amp;title=Impact+of+mycotoxins+on+the+intestine:+are+mucus+and+microbiota+new+targets?&amp;author=H+Robert&amp;author=D+Payros&amp;author=P+Pinton&amp;author=V+Theodorou&amp;author=M+Mercier-Bonin&amp;volume=20&amp;publication_year=2017&amp;pages=249-275.10.1080\/10937404&amp;pmid=28636450&amp;doi=10.1080\/10937404.2017.1326071&amp;\" target=\"_blank\" rel=\"noopener noreferrer\" ref=\"reftype=other&amp;article-id=6373143&amp;issue-id=327480&amp;journal-id=1872&amp;FROM=Article%7CCitationRef&amp;TO=Content%20Provider%7CLink%7CGoogle%20Scholar\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">Google Scholar<\/a>]<\/span><\/span><\/div>\n<\/div>\n<\/div>\n<div style=\"display: none; width: 200px; top: -100px; left: -100px;\" aria-live=\"assertive\" aria-hidden=\"true\" class=\"ui-helper-reset ui-ncbipopper-wrapper ui-ncbilinksmenu\">\n<ul id=\"ui-ncbiinpagenav-2\">\n<li><a href=\"#Abs1title\">Abstract<\/a><\/li>\n<li><a href=\"#Sec1title\">Background<\/a><\/li>\n<li><a href=\"#Sec2title\">Methods<\/a><\/li>\n<li><a href=\"#Sec13title\">Results<\/a><\/li>\n<li><a href=\"#Sec21title\">Discussion<\/a><\/li>\n<li><a href=\"#Sec22title\">Conclusions<\/a><\/li>\n<li><a href=\"#sec-a.o.ftitle\">Additional files<\/a><\/li>\n<li><a href=\"#ack-a.p.btitle\">Acknowledgements<\/a><\/li>\n<li><a href=\"#glossary-a.p.atitle\">Abbreviations<\/a><\/li>\n<li><a href=\"#notes-a.p.ctitle\">Authors\u2019 contributions<\/a><\/li>\n<li><a href=\"#notes-a.p.dtitle\">Notes<\/a><\/li>\n<li><a href=\"#FPar3title\">Ethics approval and consent to participate<\/a><\/li>\n<li><a href=\"#FPar4title\">Consent for publication<\/a><\/li>\n<li><a href=\"#FPar5title\">Competing interests<\/a><\/li>\n<li><a href=\"#article-aaff-infotitle\">Contributor Information<\/a><\/li>\n<li><a href=\"#Bib1title\">References<\/a><\/li>\n<\/ul>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Morphological and molecular response of small intestine to lactulose and hydrogen-rich water in female piglets fed Fusarium mycotoxins contaminated diet<\/p>\n","protected":false},"author":1,"featured_media":17899,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[130],"tags":[],"disease":[930],"body-organ":[1025],"applications":[680],"test_subjects":[1524],"report-topic":[1391],"class_list":["post-27017","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-hydrogen-health","disease-food-poisoning-2","body-organ-intestine-2","applications-ingestion-2","test_subjects-pig-2","report-topic-mycotoxicosis-2"],"acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v28.1 - 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