{"id":26838,"date":"2024-01-03T21:39:07","date_gmt":"2024-01-03T19:39:07","guid":{"rendered":"https:\/\/hho-bulgaria.com\/h2-gas-prevents-liver-injury-by-inhibiting-jnk-activation\/"},"modified":"2024-01-29T21:31:42","modified_gmt":"2024-01-29T19:31:42","slug":"h2-gas-prevents-liver-injury-by-inhibiting-jnk-activation","status":"publish","type":"post","link":"https:\/\/hho-bulgaria.com\/en\/h2-gas-prevents-liver-injury-by-inhibiting-jnk-activation\/","title":{"rendered":"H2 gas prevents liver injury by inhibiting JNK activation"},"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\">Exp Ther Med.<\/a><\/span> 2021 May; 21(5): 453. <\/div>\n<div class=\"part2\"><span class=\"fm-vol-iss-date\">Published online 2021 Mar 1. <\/span>  <span class=\"doi\"><span>doi:&nbsp;<\/span><a href=\"\/\/doi.org\/10.3892%2Fetm.2021.9884\" target=\"_blank\" rel=\"noopener noreferrer\" ref=\"reftype=other&amp;article-id=7976433&amp;issue-id=377760&amp;journal-id=1895&amp;FROM=Article%7CFront%20Matter&amp;TO=Content%20Provider%7CCrosslink%7CDOI\">10.3892\/etm.2021.9884<\/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>PMC7976433<\/span><\/div>\n<div class=\"fm-citation-pmid\">PMID: <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/33767761\">33767761<\/a><\/div>\n<\/div>\n<\/div>\n<h1 class=\"content-title\">Hydrogen gas alleviates acute alcohol-induced liver injury by inhibiting JNK activation<\/h1>\n<div class=\"half_rhythm\">\n<div class=\"contrib-group fm-author\"><a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Zhang%20Y%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139685800214592\" co-class=\"co-affbox\">Yaxing Zhang<\/a>,<sup>1<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Bi%20M%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139685784320688\" co-class=\"co-affbox\">Mingmin Bi<\/a>,<sup>2,<\/sup><sup>3<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Chen%20Z%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139685783708528\" co-class=\"co-affbox\">Zifeng Chen<\/a>,<sup>4,<\/sup><sup>5<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Dai%20M%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139685783706480\" co-class=\"co-affbox\">Min Dai<\/a>,<sup>1<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Zhou%20G%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139685786519376\" co-class=\"co-affbox\">Ge Zhou<\/a>,<sup>1<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Hu%20Y%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139685786517392\" co-class=\"co-affbox\">Yuxuan Hu<\/a>,<sup>1<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Yang%20H%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139685784999056\" co-class=\"co-affbox\">Hongzhi Yang<\/a>,<sup>1<\/sup> and  <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Guan%20W%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm139685784997072\" co-class=\"co-affbox\">Weibing Guan<\/a><sup>1<\/sup><sup><\/sup><\/div>\n<div style=\"display:none\" class=\"contrib-group aff-tip\">\n<div id=\"_co_idm139685800214592\">\n<h3 class=\"no_margin\">Yaxing Zhang<\/h3>\n<p><sup>1<\/sup>Department of Traditional Chinese Medicine, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Zhang%20Y%5BAuthor%5D\">Yaxing Zhang<\/a><\/div>\n<\/div>\n<div id=\"_co_idm139685784320688\">\n<h3 class=\"no_margin\">Mingmin Bi<\/h3>\n<p><sup>2<\/sup>Department of Otorhinolaryngology, The Seventh Affiliated Hospital, Sun Yat-sen University, Shenzhen, Guangdong 518107, P.R. China<\/p>\n<p><sup>3<\/sup>Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong 510080, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Bi%20M%5BAuthor%5D\">Mingmin Bi<\/a><\/div>\n<\/div>\n<div id=\"_co_idm139685783708528\">\n<h3 class=\"no_margin\">Zifeng Chen<\/h3>\n<p><sup>4<\/sup>Biofeedback Laboratory, Xinhua College of Sun Yat-sen University, Guangzhou, Guangdong 510520, P.R. China<\/p>\n<p><sup>5<\/sup>Department of Biomedical Engineering, Xinhua College of Sun Yat-sen University, Guangzhou, Guangdong 510520, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Chen%20Z%5BAuthor%5D\">Zifeng Chen<\/a><\/div>\n<\/div>\n<div id=\"_co_idm139685783706480\">\n<h3 class=\"no_margin\">Min Dai<\/h3>\n<p><sup>1<\/sup>Department of Traditional Chinese Medicine, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Dai%20M%5BAuthor%5D\">Min Dai<\/a><\/div>\n<\/div>\n<div id=\"_co_idm139685786519376\">\n<h3 class=\"no_margin\">Ge Zhou<\/h3>\n<p><sup>1<\/sup>Department of Traditional Chinese Medicine, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Zhou%20G%5BAuthor%5D\">Ge Zhou<\/a><\/div>\n<\/div>\n<div id=\"_co_idm139685786517392\">\n<h3 class=\"no_margin\">Yuxuan Hu<\/h3>\n<p><sup>1<\/sup>Department of Traditional Chinese Medicine, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Hu%20Y%5BAuthor%5D\">Yuxuan Hu<\/a><\/div>\n<\/div>\n<div id=\"_co_idm139685784999056\">\n<h3 class=\"no_margin\">Hongzhi Yang<\/h3>\n<p><sup>1<\/sup>Department of Traditional Chinese Medicine, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Yang%20H%5BAuthor%5D\">Hongzhi Yang<\/a><\/div>\n<\/div>\n<div id=\"_co_idm139685784997072\">\n<h3 class=\"no_margin\">Weibing Guan<\/h3>\n<p><sup>1<\/sup>Department of Traditional Chinese Medicine, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=Guan%20W%5BAuthor%5D\">Weibing Guan<\/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=\"idm139685785839520_ai\">Author information<\/a> <a href=\"#\" class=\"pmctoggle\" rid=\"idm139685785839520_an\">Article notes<\/a> <a href=\"#\" class=\"pmctoggle\" rid=\"idm139685785839520_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=\"idm139685785839520_ai\" style=\"display:none\">\n<div class=\"fm-affl\" id=\"af1-etm-0-0-09884\"><sup>1<\/sup>Department of Traditional Chinese Medicine, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510630, P.R. China<\/div>\n<div class=\"fm-affl\" id=\"af2-etm-0-0-09884\"><sup>2<\/sup>Department of Otorhinolaryngology, The Seventh Affiliated Hospital, Sun Yat-sen University, Shenzhen, Guangdong 518107, P.R. China<\/div>\n<div class=\"fm-affl\" id=\"af3-etm-0-0-09884\"><sup>3<\/sup>Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong 510080, P.R. China<\/div>\n<div class=\"fm-affl\" id=\"af4-etm-0-0-09884\"><sup>4<\/sup>Biofeedback Laboratory, Xinhua College of Sun Yat-sen University, Guangzhou, Guangdong 510520, P.R. China<\/div>\n<div class=\"fm-affl\" id=\"af5-etm-0-0-09884\"><sup>5<\/sup>Department of Biomedical Engineering, Xinhua College of Sun Yat-sen University, Guangzhou, Guangdong 510520, P.R. China<\/div>\n<div id=\"c1-etm-0-0-09884\"><em>Correspondence to:<\/em> Dr Weibing Guan or Professor Hongzhi Yang, Department of Traditional Chinese Medicine, The Third Affiliated Hospital, Sun Yat-sen University, 600 Tianhe Road, Guangzhou, Guangdong 510630, P.R. China <a href=\"mailto:dev@null\" data-email=\"nc.ude.usys.liam@bwnaug\" class=\"oemail\">nc.ude.usys.liam@bwnaug<\/a><span style=\"unicode-bidi:bidi-override\">, <\/span><a href=\"mailto:dev@null\" data-email=\"nc.ude.usys.liam@ihzhgnay\" class=\"oemail\">nc.ude.usys.liam@ihzhgnay<\/a&gt;; <a href=\"mailto:dev@null\" data-email=\"moc.361@0691gnayzh\" class=\"oemail\">moc.361@0691gnayzh<\/a><\/div>\n<\/div>\n<div class=\"fm-article-notes hide half_rhythm\" id=\"idm139685785839520_an\" style=\"display:none\">\n<div class=\"fm-pubdate half_rhythm\">Received 2020 Jun 21; Accepted 2020 Dec 8.<\/div>\n<\/div>\n<div class=\"permissions half_rhythm hide\" id=\"idm139685785839520_cpl\" style=\"display:none\">\n<div class=\"fm-copyright half_rhythm\"><a href=\"\/pmc\/about\/copyright\/\">Copyright<\/a> : \u00a9 Zhang et al.<\/div>\n<div class=\"license half_rhythm\">This is an open access article distributed under the terms of the <a href=\"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/\" data-ga-action=\"click_feat_suppl\" ref=\"reftype=extlink&amp;article-id=7976433&amp;issue-id=377760&amp;journal-id=1895&amp;FROM=Article%7CFront%20Matter&amp;TO=External%7CLink%7CURI\" target=\"_blank\" rel=\"noopener\">Creative Commons Attribution-NonCommercial-NoDerivs License<\/a>, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.<\/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-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 analyzed during the current study are available from the corresponding author on reasonable request.<\/p>\n<\/dd>\n<\/dl>\n<\/div>\n<div id=\"abstract-a.g.b.t\" 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=\"abstract-a.g.b.ttitle\">Abstract<\/h2>\n<p><!--article-meta--><\/p>\n<div>\n<p class=\"p p-first-last\">Binge alcohol drinking is fast becoming a global health concern, with the liver among the first organ involved and the one afflicted with the greatest degree of injury. Oxidative stress, alterations in hepatic metabolism, immunity and inflammation have all been reported to contribute to the development of alcoholic liver disease (ALD). Hydrogen gas (H<sub>2<\/sub>) serves a key role in the modulation of hepatic redox, immune and inflammatory homeostasis. However, the effects of treatment using intraperitoneal injection of H<sub>2<\/sub> on ALD remain unexplored. Therefore, the aim of the present study was to investigate the effects and underlying mechanism of intraperitoneal injection of H<sub>2<\/sub> on acute alcohol-induced liver injury in a mouse model. H<sub>2<\/sub> was administered by daily intraperitoneal injections (1.0 ml\/100 g) for 4 days. On day 4, the mice received H<sub>2<\/sub> after fasting for 5.5 h. After 30 min, the mice were administered with 33% (v\/v) ethanol at a cumulative dose of 4.5 g\/kg body weight by four equally divided gavages at 20-min intervals. Blood and liver tissues were collected at 16 h after the first ethanol gavage. Subsequently, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride and total cholesterol (TC) levels were analyzed using an Automatic Clinical Analyzer. Hepatic JNK activity and GAPDH levels were examined by western blotting. It was observed that acute ethanol gavage induced liver injury, as indicated by significantly increased serum ALT and AST levels, which were effectively decreased by H<sub>2<\/sub> at 16 h after the first ethanol gavage. In addition, H<sub>2<\/sub> treatment reduced serum TC levels in the Alcohol+H<sub>2<\/sub> group when compared with those in Alcohol group. Mechanistically, H<sub>2<\/sub> attenuated hepatic JNK phosphorylation induced by acute ethanol gavage. Therefore, the results of the present study demonstrated that treatment with exogenous H<sub>2<\/sub> by intraperitoneal injection may alleviate acute alcohol-induced liver injury by inhibiting hepatic JNK activation, which may represent a novel therapeutic strategy for ALD.<\/p>\n<\/div>\n<div class=\"sec\"><strong class=\"kwd-title\">Keywords: <\/strong><span class=\"kwd-text\">hydrogen, alcohol, alanine aminotransferase, aspartate aminotransferase, total cholesterol, triglyceride, c-Jun N-terminal kinase<\/span><\/div>\n<\/div>\n<div id=\"sec-a.h.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=\"sec-a.h.atitle\">Introduction<\/h2>\n<p class=\"p p-first\">Excessive alcohol intake, as result of binge drinking as well as chronic alcohol consumption of &gt;40 g per day, is becoming a global healthcare concern (<a href=\"#b1-etm-0-0-09884\" rid=\"b1-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">1<\/a>,<a href=\"#b2-etm-0-0-09884\" rid=\"b2-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">2<\/a>). Globally, ~2 billion individuals consume alcohol regularly, where &gt;75 million are diagnosed with disorders associated with alcohol abuse and are at risk of developing alcohol-associated liver diseases (<a href=\"#b3-etm-0-0-09884\" rid=\"b3-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">3<\/a>). The liver typically sustains the earliest and the greatest degree of tissue injury caused by excessive drinking, since it is the primary site of ethanol metabolism (<a href=\"#b1-etm-0-0-09884\" rid=\"b1-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">1<\/a>,<a href=\"#b4-etm-0-0-09884\" rid=\"b4-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">4<\/a>). Excessive alcohol consumption causes alcoholic liver disease (ALD), which is characterized by a wide spectrum of hepatic pathologies, from reversible fatty liver (simple steatosis) to acute alcoholic hepatitis, chronic fibrosis and cirrhosis and superimposed hepatocellular carcinoma (HCC) (<a href=\"#b2-etm-0-0-09884\" rid=\"b2-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">2<\/a>,<a href=\"#b5-etm-0-0-09884\" rid=\"b5-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">5<\/a>,<a href=\"#b6-etm-0-0-09884\" rid=\"b6-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">6<\/a>). Mechanistically, pathogenesis of ALD includes genetic susceptibility, oxidative stress, hepatic metabolism alteration and steatosis, acetaldehyde-mediated toxicity, cytokine- and chemokine-induced inflammation, alterations in immunity and dysbiosis, epigenetic changes and modifications in the regenerative process (<a href=\"#b2-etm-0-0-09884\" rid=\"b2-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">2<\/a>,<a href=\"#b6-etm-0-0-09884\" rid=\"b6-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">6-8<\/a>). Alcohol abstinence achieved by psychosomatic intervention is at present the best treatment for all stages of ALD (<a href=\"#b2-etm-0-0-09884\" rid=\"b2-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">2<\/a>). However, drinking discontinuation may be difficult, for example, the cheap price of hard liquor, easy accessibility to alcohol, and alcohol advertisement make it very difficult to prevent the increase in ALD (<a href=\"#b9-etm-0-0-09884\" rid=\"b9-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">9<\/a>). Therefore, new strategies for alleviating alcohol-induced liver injury remain to be in demand.<\/p>\n<p>Hydrogen gas (H<sub>2<\/sub>) is the lightest and diffusible gas molecule that has been shown to confer potent antioxidant and anti-inflammatory properties (<a href=\"#b10-etm-0-0-09884\" rid=\"b10-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">10<\/a>,<a href=\"#b11-etm-0-0-09884\" rid=\"b11-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">11<\/a>). It can be absorbed into the blood circulation, such that it reaches the target organ either by blood circulation or free diffusion (<a href=\"#b11-etm-0-0-09884\" rid=\"b11-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">11<\/a>). Therefore, treatments involving exogenous H<sub>2<\/sub>, including breathing H<sub>2<\/sub> gas, injection with H<sub>2<\/sub>-rich saline and drinking H<sub>2<\/sub>-rich water, may protect against excessive oxidative stress- and inflammation-related liver damage, including liver injury induced by drugs, sepsis, bile duct ligation, ischemia\/reperfusion (I\/R), CO<sub>2<\/sub> pneumoperitoneum and chronic intermittent hypoxia, in addition to non-alcoholic fatty liver disease (NAFLD) (<a href=\"#b12-etm-0-0-09884\" rid=\"b12-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">12-17<\/a>). Furthermore, drinking H<sub>2<\/sub>-rich water has been indicated to protect against chronic ethanol-induced hepatotoxicity (<a href=\"#b18-etm-0-0-09884\" rid=\"b18-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">18<\/a>). These previous observations suggest that H<sub>2<\/sub> serves an important role in modulating hepatic redox, immune and inflammatory homeostasis (<a href=\"#b11-etm-0-0-09884\" rid=\"b11-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">11<\/a>).<\/p>\n<p>Previous studies have demonstrated that supplementation with exogenous H<sub>2<\/sub> by intraperitoneal injection may improve lipopolysaccharide (LPS)-induced cardiac dysfunction, isoproterenol-induced cardiac hypertrophy, pressure overload-induced vascular hypertrophy, and cardiopulmonary cerebral resuscitation in a cardiac arrest rabbit model (<a href=\"#b19-etm-0-0-09884\" rid=\"b19-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">19-23<\/a>). These were mediated by the suppression of excessive oxidative stress and inflammatory responses (<a href=\"#b19-etm-0-0-09884\" rid=\"b19-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">19-23<\/a>). However, the effects of intraperitoneal injection of H<sub>2<\/sub> on ALD remain unclear.<\/p>\n<p class=\"p p-last\">Therefore, the aim of the present study was to investigate the effect of H<sub>2<\/sub> intraperitoneal injection on acute alcohol-induced liver injury in a mouse model and to elucidate the potential underlying mechanisms.<\/p>\n<\/div>\n<div id=\"sec-a.h.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=\"sec-a.h.btitle\">Materials and methods<\/h2>\n<div id=\"sec-a.h.b.b\" class=\"sec sec-first-last\">\n<div id=\"sec-a.h.b.b.b\" class=\"sec sec-first\">\n<h4 id=\"sec-a.h.b.b.btitle\" class=\"inline\">Drugs <\/h4>\n<p class=\"p p-first-last\">In total, 99,999% H<sub>2<\/sub> (Dalian Special Gases Co., Ltd.) was injected into a vacuumed aseptic soft plastic infusion bag (100 ml; Hebei Tiancheng Pharmaceutical Co., Ltd.) from a seamless steel gas cylinder under sterile conditions, as previously described (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f1-etm-0-0-09884\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f1-etm-0-0-09884\" rid-ob=\"ob-f1-etm-0-0-09884\" co-legend-rid=\"lgnd_f1-etm-0-0-09884\" rel=\"noopener\"><span>Fig. 1<\/span><\/a>) (<a href=\"#b20-etm-0-0-09884\" rid=\"b20-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20-23<\/a>). Anhydrous ethanol (Guangzhou Chemical Reagent Factory) was dissolved in double-distilled water to obtain 33% (v\/v) ethanol (0.26 g\/ml) (<a href=\"#b24-etm-0-0-09884\" rid=\"b24-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>).<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"f1-etm-0-0-09884\" co-legend-rid=\"lgnd_f1-etm-0-0-09884\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f1-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f1-etm-0-0-09884\" rid-ob=\"ob-f1-etm-0-0-09884\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139685793629872\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f1-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f1-etm-0-0-09884\" rid-ob=\"ob-f1-etm-0-0-09884\" 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=7976433_etm-21-05-09884-g00.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 etm-21-05-09884-g00.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/etm-21-05-09884-g00.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139685793629872\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f1-etm-0-0-09884\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_f1-etm-0-0-09884\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f1-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f1-etm-0-0-09884\" rid-ob=\"ob-f1-etm-0-0-09884\" rel=\"noopener\">Figure 1<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Preparation of H<sub>2<\/sub> in the storage container. (A) Sterile saline and the gases in the aseptic soft plastic infusion bag (100 ml) were emptied using a 50-ml syringe under sterile conditions. (B) The gas outlet of seamless steel gas cylinder was connected to a silica gel tube, which was in turn connected to a 1-ml syringe needle at the other end. This syringe needle was inserted into the vacuumed plastic bag, which was filled by opening the reducing valve of the H<sub>2<\/sub> steel cylinder until the bag was full of H<sub>2<\/sub> gas with no dead volume.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.h.b.b.c\" class=\"sec\">\n<h4 id=\"sec-a.h.b.b.ctitle\" class=\"inline\">Animal model of acute alcohol-induced liver injury and treatment protocol <\/h4>\n<p class=\"p p-first\">Male C57BL\/6J mice (Animal license no. 44007200061823) were purchased from the Guangdong Medical Laboratory Animal Center (Foshan, China). A total of 28 mice aged 8-10 weeks were used in this study. All animals were housed in a temperature-controlled animal facility (21-24\u02daC) with a 12-hour light-dark cycle, and the animals had access to rodent chow and water <em>ad libitum<\/em> (<a href=\"#b20-etm-0-0-09884\" rid=\"b20-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>). All mice were provided with humane care according to the Principles of Laboratory Animal Care formulated by the National Society of Medical Research and the Guide for the Care and Use of Laboratory Animals published by the NIH (8<sup>th<\/sup> Edition, Revised 2011) (<a href=\"#b20-etm-0-0-09884\" rid=\"b20-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>,<a href=\"#b25-etm-0-0-09884\" rid=\"b25-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">25<\/a>). All animal procedures were approved by the Institutional Animal Care and Use Committee of Zhongshan School of Medicine, Sun Yat-sen University (Guangzhou, China).<\/p>\n<p class=\"p p-last\">Mice were randomly assigned into the following four groups (n=7 mice per group): i) Control; ii) Alcohol; iii) Alcohol+H<sub>2<\/sub&gt;; and iv) H<sub>2<\/sub>. In the Alcohol+H<sub>2<\/sub> and H<sub>2<\/sub> groups, H<sub>2<\/sub> was administered daily at the dose of 1.0 ml\/100 g by intraperitoneal injection for 4 days. On day 4, mice in each group were fasted for 6 h before the mice in the Alcohol and Alcohol+H<sub>2<\/sub> groups were orally administered with 33% (v\/v) ethanol at a cumulative dose of 4.5 g\/kg body weight (17.3 ml\/kg body weight) by four equally divided gavages at 20-min intervals (<a href=\"#b24-etm-0-0-09884\" rid=\"b24-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>). Mice in the Alcohol+H<sub>2<\/sub> and H<sub>2<\/sub> groups were administered with an intraperitoneal injection (1.0 ml\/100 g) of H<sub>2<\/sub> 30 min before the first ethanol administration (<a href=\"#b20-etm-0-0-09884\" rid=\"b20-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>). In addition, mice in the Control and H<sub>2<\/sub> groups received the same volume of double-distilled water (17.3 ml\/kg body weight) by four equally divided gavages at 20 min intervals. The animals were anesthetized by intraperitoneal injection of ketamine (100 mg\/kg) and xylazine (10 mg\/kg) after 16 h of the first ethanol gavage, cardiac puncture was performed when the animals reached the surgical plane of anesthesia and ~0.4-0.5 ml blood was collected from each mouse (<a href=\"#b26-etm-0-0-09884\" rid=\"b26-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">26-28<\/a>). Euthanasia was then performed by cervical dislocation in a state of deep anesthesia before liver tissues were extracted (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f2-etm-0-0-09884\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f2-etm-0-0-09884\" rid-ob=\"ob-f2-etm-0-0-09884\" co-legend-rid=\"lgnd_f2-etm-0-0-09884\" rel=\"noopener\"><span>Fig. 2<\/span><\/a>) (<a href=\"#b24-etm-0-0-09884\" rid=\"b24-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>). The body weight of animals in each group was recorded at the baseline (Control, 25.19\u00b11.58 g; Alcohol, 24.63\u00b11.42 g; Alcohol+H<sub>2<\/sub>, 25.17\u00b11.15 g; H<sub>2<\/sub>, 23.9\u00b10.70 g) and after intervention (Control, 24.90\u00b11.36 g; Alcohol, 25.16\u00b11.53 g; Alcohol+H<sub>2<\/sub>, 25.00\u00b10.93 g; H<sub>2<\/sub>, 23.71\u00b10.63 g).<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"f2-etm-0-0-09884\" co-legend-rid=\"lgnd_f2-etm-0-0-09884\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f2-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f2-etm-0-0-09884\" rid-ob=\"ob-f2-etm-0-0-09884\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139685802001632\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f2-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f2-etm-0-0-09884\" rid-ob=\"ob-f2-etm-0-0-09884\" 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=7976433_etm-21-05-09884-g01.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 etm-21-05-09884-g01.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/etm-21-05-09884-g01.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139685802001632\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f2-etm-0-0-09884\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_f2-etm-0-0-09884\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f2-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f2-etm-0-0-09884\" rid-ob=\"ob-f2-etm-0-0-09884\" rel=\"noopener\">Figure 2<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Experimental procedures. On days 1-3, mice in the Alcohol+H<sub>2<\/sub> and H<sub>2<\/sub> groups were intraperitoneally injected with H<sub>2<\/sub> (1.0 ml\/100 g) at 18:00 daily. On day 4, the mice were fasted starting at 12:00 for 6 h before the first ethanol administration. Mice in the Alcohol+H<sub>2<\/sub> and H<sub>2<\/sub> groups received H<sub>2<\/sub> at 17:30. Mice in the Alcohol and Alcohol+H<sub>2<\/sub> groups were orally administered 33% (v\/v) ethanol at an accumulative dosage of 4.5 g\/kg body weight using four equally divided gavages at 18:00, 18:20, 18:40 and 19:00. Mice in the Control and H<sub>2<\/sub> groups received the same volume of ddH<sub>2<\/sub>O. On Day 5, the mice were anesthetized and euthanized for blood and tissue collection at 10:00.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.h.b.b.d\" class=\"sec\">\n<h4 id=\"sec-a.h.b.b.dtitle\" class=\"inline\">Biochemical analysis <\/h4>\n<p class=\"p p-first-last\">Blood biochemical analysis was performed as previously described (<a href=\"#b29-etm-0-0-09884\" rid=\"b29-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">29<\/a>). Briefly, the blood samples were centrifuged at the speed of 986 x g for 10 min at 4\u02daC to separate the serum. Levels of liver function markers alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in addition to the levels of lipid markers triglyceride (TG) and total cholesterol (TC) in the serum were analyzed using an Automatic Clinical Analyzer (Hitachi 7600; Hitachi High-Technologies Corporation) at the Department of Clinical Laboratory, The Third Affiliated Hospital of Sun Yat-sen University.<\/p>\n<\/div>\n<div id=\"sec-a.h.b.b.e\" class=\"sec\">\n<h4 id=\"sec-a.h.b.b.etitle\" class=\"inline\">Western blotting <\/h4>\n<p class=\"p p-first\">JNK antibody (cat. no. 9252S), phosphorylated (p-) JNK antibody (cat. no. 9255S), anti-rabbit IgG horseradish peroxidase (HRP)-conjugated secondary antibody (cat. no. 7074S) and anti-mouse IgG HRP-conjugated secondary antibody (cat. no. 7076S) were purchased from Cell Signaling Technology, Inc. GAPDH antibody (cat. no. MB001) was obtained from Bioworld Technology, Inc. BSA (cat. no. ST023-200g) was purchased from Beyotime Institute of Biotechnology.<\/p>\n<p class=\"p p-last\">Western blotting was performed as previously described (<a href=\"#b30-etm-0-0-09884\" rid=\"b30-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">30<\/a>). The proteins were transferred onto polyvinylidene fluoride membranes (EMD Millipore). Membranes were incubated in blocking buffer (1X TBST with 5% BSA) for 30 min at room temperature. For antibody incubations, membranes were incubated with primary antibodies in antibody dilution buffer (1X TBST with 5% BSA) with gentle agitation overnight at 4\u02daC (p-JNK and JNK: 1:2,000; GAPDH: 1:10,000), and with secondary antibodies in antibody dilution buffer (1X TBST with 5% BSA) with gentle agitation for 1 h at room temperature (anti-rabbit IgG HRP-conjugated secondary antibody and anti-mouse IgG HRP-conjugated secondary antibody: 1:2,000). Immobilon\u2122 Western Chemiluminescent HRP Substrate (ECL; cat. no. WBKLS0100, EMD Millipore) was used to reveal the bands using the ChemiDoc\u2122 Touch Imaging System (Bio-Rad Laboratories, Inc.). Image J software (Version 1.52; National Institutes of Health) was used for estimating the \u2018IntDen\u2019 value of the western blot bands for quantification.<\/p>\n<\/div>\n<div id=\"sec-a.h.b.b.f\" class=\"sec sec-last\">\n<h4 id=\"sec-a.h.b.b.ftitle\" class=\"inline\">Statistical analysis <\/h4>\n<p class=\"p p-first-last\">All statistical analyses were performed using GraphPad Prism 5.0 (GraphPad Software, Inc.). Data were expressed as the mean \u00b1 SD. For biochemical analysis, n=7 mice in each group. For western blotting, n=5 mice in each group. Statistical analysis was performed by one-way ANOVA followed by Bonferroni&#8217;s post hoc test. P&lt;0.05 was considered to indicate statistically significant differences.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.h.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=\"sec-a.h.ctitle\">Results<\/h2>\n<div id=\"sec-a.h.c.b\" class=\"sec sec-first-last\">\n<div id=\"sec-a.h.c.b.b\" class=\"sec sec-first\">\n<h4 id=\"sec-a.h.c.b.btitle\" class=\"inline\">H<sub>2<\/sub> treatment prevents acute alcohol-induced liver damage <\/h4>\n<p class=\"p p-first-last\">To investigate the effects of H<sub>2<\/sub> on alcohol-induced liver injury, an acute alcohol-induced hepatotoxicity mouse model was established as previously described (<a href=\"#b24-etm-0-0-09884\" rid=\"b24-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>). This model was shown to closely mimic excessive ethanol consumption in humans in terms of blood alcohol levels, behavioral and physiological effects (<a href=\"#b24-etm-0-0-09884\" rid=\"b24-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>). Compared with those in the Control group, serum ALT and AST levels in the Alcohol group were significantly higher following acute alcohol gavage (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f3-etm-0-0-09884\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f3-etm-0-0-09884\" rid-ob=\"ob-f3-etm-0-0-09884\" co-legend-rid=\"lgnd_f3-etm-0-0-09884\" rel=\"noopener\"><span>Fig. 3<\/span><\/a>), whilst intraperitoneal injection of H<sub>2<\/sub> significantly prevented these elevations in serum ALT and AST levels in mice following alcohol gavage (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f3-etm-0-0-09884\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f3-etm-0-0-09884\" rid-ob=\"ob-f3-etm-0-0-09884\" co-legend-rid=\"lgnd_f3-etm-0-0-09884\" rel=\"noopener\"><span>Fig. 3<\/span><\/a>). However, H<sub>2<\/sub> alone exerted no effects on serum ALT and AST levels compared with those in the Control group (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f3-etm-0-0-09884\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f3-etm-0-0-09884\" rid-ob=\"ob-f3-etm-0-0-09884\" co-legend-rid=\"lgnd_f3-etm-0-0-09884\" rel=\"noopener\"><span>Fig. 3<\/span><\/a>). Therefore, these observations suggest that intraperitoneal injection of H<sub>2<\/sub> effectively alleviated acute alcohol-induced liver injury in mice.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"f3-etm-0-0-09884\" co-legend-rid=\"lgnd_f3-etm-0-0-09884\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f3-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f3-etm-0-0-09884\" rid-ob=\"ob-f3-etm-0-0-09884\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139685801241712\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f3-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f3-etm-0-0-09884\" rid-ob=\"ob-f3-etm-0-0-09884\" 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=7976433_etm-21-05-09884-g02.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 etm-21-05-09884-g02.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/etm-21-05-09884-g02.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139685801241712\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f3-etm-0-0-09884\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_f3-etm-0-0-09884\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f3-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f3-etm-0-0-09884\" rid-ob=\"ob-f3-etm-0-0-09884\" rel=\"noopener\">Figure 3<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>H<sub>2<\/sub> treatment reduces serum ALT and AST levels after ethanol feeding. Serum (A) ALT and (B) AST levels. n=7 mice in each group. <sup>***<\/sup>P&lt;0.001 vs. Control and vs. H<sub>2<\/sub&gt;; <sup>#<\/sup>P&lt;0.05 vs. Control and vs. Alcohol; <sup>##<\/sup>P&lt;0.01 vs. Alcohol; <sup>&amp;&amp;<\/sup>P&lt;0.01 vs. Alcohol+H<sub>2<\/sub>. ALT, alanine aminotransferase; AST, aspartate aminotransferase.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.h.c.b.c\" class=\"sec\">\n<h4 id=\"sec-a.h.c.b.ctitle\" class=\"inline\">Effects of H<sub>2<\/sub> on serum lipid levels in acute alcohol-induced liver injury <\/h4>\n<p class=\"p p-first-last\">Long-term chronic alcohol consumption may lead to disruptions in lipid metabolism (<a href=\"#b6-etm-0-0-09884\" rid=\"b6-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">6<\/a>). Therefore, the serum levels of TC and TG were next examined. Although acute alcohol treatment slightly increased serum TC levels, there was no significant difference between those in the Control and Alcohol groups (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f4-etm-0-0-09884\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f4-etm-0-0-09884\" rid-ob=\"ob-f4-etm-0-0-09884\" co-legend-rid=\"lgnd_f4-etm-0-0-09884\" rel=\"noopener\"><span>Fig. 4A<\/span><\/a>). Serum TC levels in the Alcohol+H<sub>2<\/sub> group were significantly lower compared with those in the Alcohol group (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f4-etm-0-0-09884\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f4-etm-0-0-09884\" rid-ob=\"ob-f4-etm-0-0-09884\" co-legend-rid=\"lgnd_f4-etm-0-0-09884\" rel=\"noopener\"><span>Fig. 4A<\/span><\/a>). However, there were no significant differences in TG levels among the four groups (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f4-etm-0-0-09884\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f4-etm-0-0-09884\" rid-ob=\"ob-f4-etm-0-0-09884\" co-legend-rid=\"lgnd_f4-etm-0-0-09884\" rel=\"noopener\"><span>Fig. 4B<\/span><\/a>). These data suggest that intraperitoneal injection of H<sub>2<\/sub> may modulate blood TC levels in an acute alcohol-induced liver injury mice model.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"f4-etm-0-0-09884\" co-legend-rid=\"lgnd_f4-etm-0-0-09884\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f4-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f4-etm-0-0-09884\" rid-ob=\"ob-f4-etm-0-0-09884\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139685791560240\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f4-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f4-etm-0-0-09884\" rid-ob=\"ob-f4-etm-0-0-09884\" 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=7976433_etm-21-05-09884-g03.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 etm-21-05-09884-g03.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/etm-21-05-09884-g03.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139685791560240\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f4-etm-0-0-09884\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_f4-etm-0-0-09884\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f4-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f4-etm-0-0-09884\" rid-ob=\"ob-f4-etm-0-0-09884\" rel=\"noopener\">Figure 4<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of H<sub>2<\/sub> on serum TC and TG levels after ethanol feeding. Serum (A) TC and (B) TG levels. n=7 mice in each group. <sup>##<\/sup>P&lt;0.01 vs. Alcohol. TC, total cholesterol; TG, triglyceride.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.h.c.b.d\" class=\"sec sec-last\">\n<h4 id=\"sec-a.h.c.b.dtitle\" class=\"inline\">H<sub>2<\/sub> alleviates acute alcohol-induced hepatic JNK activation <\/h4>\n<p class=\"p p-first-last\">Acute ethanol loading causes oxidative stress and activates cell-death signaling through the JNK pathway in the liver (<a href=\"#b31-etm-0-0-09884\" rid=\"b31-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">31<\/a>). In addition, JNK is a key mediator in hepatic steatosis, where it regulates transcription factor activity associated with lipid metabolism (<a href=\"#b32-etm-0-0-09884\" rid=\"b32-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">32<\/a>,<a href=\"#b33-etm-0-0-09884\" rid=\"b33-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">33<\/a>). In the present study, the upregulation of serum ALT and AST levels after ethanol treatment indicated hepatocytes damage. Therefore, the role of hepatic JNK activation in the potentially protective effects of H<sub>2<\/sub> against acute alcohol-induced liver injury was examined. Western blotting data demonstrated that acute ethanol treatment significantly increased hepatic JNK phosphorylation, which was significantly prevented by intraperitoneal injection with H<sub>2<\/sub> (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f5-etm-0-0-09884\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f5-etm-0-0-09884\" rid-ob=\"ob-f5-etm-0-0-09884\" co-legend-rid=\"lgnd_f5-etm-0-0-09884\" rel=\"noopener\"><span>Fig. 5<\/span><\/a>). Therefore, these data suggest that intraperitoneal injection with H<sub>2<\/sub> conferred protective effects against acute alcohol-induced liver injury by preventing hepatic JNK activation.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"f5-etm-0-0-09884\" co-legend-rid=\"lgnd_f5-etm-0-0-09884\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f5-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f5-etm-0-0-09884\" rid-ob=\"ob-f5-etm-0-0-09884\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm139685791555696\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f5-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f5-etm-0-0-09884\" rid-ob=\"ob-f5-etm-0-0-09884\" 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=7976433_etm-21-05-09884-g04.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 etm-21-05-09884-g04.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/etm-21-05-09884-g04.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm139685791555696\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f5-etm-0-0-09884\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_f5-etm-0-0-09884\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC7976433\/figure\/f5-etm-0-0-09884\/\" target=\"figure\" rid-figpopup=\"f5-etm-0-0-09884\" rid-ob=\"ob-f5-etm-0-0-09884\" rel=\"noopener\">Figure 5<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects on H<sub>2<\/sub> on hepatic JNK activation after ethanol feeding. (A) Representative western blotting images of p-JNK and JNK and GAPDH. (B) Quantification of p-JNK\/total JNK ratio. The averages of the p-JNK\/JNK ratio in the Control group were set as \u20181\u2019, and all values were normalized to \u20181\u2019. n=5 mice in each group. <sup>**<\/sup>P&lt;0.01 vs. Control; <sup>#<\/sup>P&lt;0.05 and <sup>###<\/sup>P&lt;0.001 vs. Alcohol.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.h.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=\"sec-a.h.dtitle\">Discussion<\/h2>\n<p class=\"p p-first\">Inflammatory and cytokine signaling, oxidative stress and mitochondrial dysfunction, alterations in hepatic metabolism and hepatocyte cell death, and abnormalities in immunity and dysbiosis are all key to the pathogenesis of liver diseases, including ALD and NAFLD (<a href=\"#b2-etm-0-0-09884\" rid=\"b2-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">2<\/a>,<a href=\"#b34-etm-0-0-09884\" rid=\"b34-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">34-36<\/a>). Therefore, strategies preventing fatty liver disease progression were previously developed using combinations of naturally occurring compounds products in animal models. For example, the combination of docosahexaenoic acid and the antioxidant hydroxytyrosol was demonstrated to prevent high-fat diet-induced liver steatosis, by inhibiting oxidative stress, mitochondrial dysfunction and inflammation associated with steatosis (<a href=\"#b34-etm-0-0-09884\" rid=\"b34-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">34<\/a>,<a href=\"#b35-etm-0-0-09884\" rid=\"b35-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">35<\/a>). H<sub>2<\/sub> was first reported to alleviate skin tumors by neutralizing toxic free radicals in 1975(<a href=\"#b37-etm-0-0-09884\" rid=\"b37-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">37<\/a>). In 2007, H<sub>2<\/sub> was demonstrated to act as a therapeutic antioxidant by selectively reducing cytotoxic hydroxyl radical levels to improve focal I\/R-induced brain injury in rats (<a href=\"#b38-etm-0-0-09884\" rid=\"b38-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">38<\/a>). Since then, H<sub>2<\/sub> has been extensively investigated, where it has been shown to be an able antioxidant, anti-inflammatory and anti-apoptotic agent (<a href=\"#b10-etm-0-0-09884\" rid=\"b10-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">10<\/a>). The present study was undertaken to determine the effect of supplementation with exogenous H<sub>2<\/sub> on acute alcohol-induced liver injury in mice.<\/p>\n<p>Exogenous H<sub>2<\/sub> can be supplied by 2% H<sub>2<\/sub> gas inhalation (<a href=\"#b38-etm-0-0-09884\" rid=\"b38-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">38<\/a>), drinking H<sub>2<\/sub>-rich water (<a href=\"#b39-etm-0-0-09884\" rid=\"b39-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">39<\/a>), intraperitoneal injection of H<sub>2<\/sub>-rich saline (<a href=\"#b12-etm-0-0-09884\" rid=\"b12-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">12<\/a>) and intraperitoneal injection of H<sub>2<\/sub> gas (<a href=\"#b19-etm-0-0-09884\" rid=\"b19-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">19<\/a>). Previous studies found that intraperitoneal injection of H<sub>2<\/sub> gas can alleviate vascular remodeling (<a href=\"#b23-etm-0-0-09884\" rid=\"b23-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">23<\/a>), cardiac hypertrophy and dysfunction (<a href=\"#b20-etm-0-0-09884\" rid=\"b20-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>,<a href=\"#b21-etm-0-0-09884\" rid=\"b21-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">21<\/a>), and display neuroprotective effects in rabbits experiencing cardiac arrest (<a href=\"#b19-etm-0-0-09884\" rid=\"b19-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">19<\/a>). In the present study, it was observed that ethanol consumption induced hepatocyte injury as indicated by the upregulation of serum ALT and AST levels. Intraperitoneal injection with H<sub>2<\/sub> gas was found to effectively protect against acute alcohol-induced liver injury and reduced serum TC levels. However, body weight was not influenced by the acute intraperitoneal injection of H<sub>2<\/sub> gas or acute alcohol feeding. The lack of food intake records, liver weight, liver histological analysis and steatosis score and hepatic TG analysis are limitations of the present study. It has been documented that chronic alcohol over consumption may lead to hepatic steatosis, fibrosis and cirrhosis and eventually HCC (<a href=\"#b2-etm-0-0-09884\" rid=\"b2-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">2<\/a>,<a href=\"#b5-etm-0-0-09884\" rid=\"b5-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">5<\/a>,<a href=\"#b6-etm-0-0-09884\" rid=\"b6-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">6<\/a>). The long-term effects of intraperitoneal H<sub>2<\/sub> injection on pathological features, such as hepatic fibrosis, and the effects of other H<sub>2<\/sub> delivery methods, such as drinking H<sub>2<\/sub>-rich alcohol, on ALD, require further study.<\/p>\n<p>Acute alcohol feeding activates cytochrome P450 2E1 and causes oxidative stress to activate JNK in hepatocytes, and JNK, in turn, reciprocally increases oxidative stress (<a href=\"#b33-etm-0-0-09884\" rid=\"b33-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">33<\/a>). JNK activation can cause programmed cell death, and increases the expression of lipogenic transcription factor sterol regulatory element binding protein (SREBP)-activated lipid synthesis enzymes, resulting in hepatic steatosis (<a href=\"#b31-etm-0-0-09884\" rid=\"b31-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">31<\/a>,<a href=\"#b33-etm-0-0-09884\" rid=\"b33-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">33<\/a>,<a href=\"#b40-etm-0-0-09884\" rid=\"b40-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">40<\/a>,<a href=\"#b41-etm-0-0-09884\" rid=\"b41-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">41<\/a>). Additionally, JNK may suppress hepatic peroxisome proliferator-activated receptor-\u03b1 (PPAR-\u03b1) activation, which is a transcription factor and a positive regulator of intracellular free fatty acid and TG metabolism by regulating gene transcription involved in fatty acid transport and degradation in mitochondria and peroxisomes (<a href=\"#b32-etm-0-0-09884\" rid=\"b32-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">32<\/a>). Therefore, hepatic JNK activation increases oxidative stress, leads to hepatocyte apoptosis and injury, and contributes to hepatic steatosis via modulating lipid metabolic transcription factor activation. H<sub>2<\/sub> has been shown to inhibit JNK activation in numerous liver disease and cardiovascular disease animal models (<a href=\"#b11-etm-0-0-09884\" rid=\"b11-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">11<\/a>,<a href=\"#b20-etm-0-0-09884\" rid=\"b20-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>,<a href=\"#b22-etm-0-0-09884\" rid=\"b22-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>,<a href=\"#b23-etm-0-0-09884\" rid=\"b23-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">23<\/a>). JNK inhibitor can improve acute alcohol-induced fatty liver and oxidative stress in mice (<a href=\"#b33-etm-0-0-09884\" rid=\"b33-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">33<\/a>). In the present study, phosphorylation of JNK in the liver induced by acute alcohol consumption was inhibited by the intraperitoneal injection of H<sub>2<\/sub>. Therefore, the protective effect of H<sub>2<\/sub> against acute alcohol-induced liver injury is hypothesized to be partially mediated by reducing JNK phosphorylation. In addition to the inhibition of JNK activation, H<sub>2<\/sub> has been shown to attenuate the activation of NF-\u03baB in an LPS-induced cardiac dysfunction mice model (<a href=\"#b20-etm-0-0-09884\" rid=\"b20-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>). H<sub>2<\/sub> has also been shown to increase the hepatic expression of nuclear factor erythroid 2-related factor 2 (Nrf2) (<a href=\"#b42-etm-0-0-09884\" rid=\"b42-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">42<\/a>) and PPAR-\u03b1 (<a href=\"#b43-etm-0-0-09884\" rid=\"b43-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">43<\/a>) and reduced the hepatic expression of SREBP-1c (<a href=\"#b44-etm-0-0-09884\" rid=\"b44-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">44<\/a>) in NAFLD animal models. These transcription factors are essential mediators of inflammation (NF-\u03baB), oxidative stress (Nrf2) and lipid metabolism (SREBP-1c and PPAR-\u03b1), where they have been reported to serve key roles in the pathogenesis of both ALD and NAFLD (<a href=\"#b35-etm-0-0-09884\" rid=\"b35-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">35<\/a>). Therefore, strategies modulating the expression or activation of these transcription factors may alleviate ALD and NAFLD (<a href=\"#b35-etm-0-0-09884\" rid=\"b35-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">35<\/a>,<a href=\"#b45-etm-0-0-09884\" rid=\"b45-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">45-51<\/a>). However, whether the protective effects of intraperitoneal injection of H<sup>2<\/sup> against acute alcohol-induced liver injury is mediated by regulating the expression and\/or activation of these transcription factors aforementioned requires further investigation.<\/p>\n<p>The gut microbiome serves an important role in liver homeostasis, intestinal dysbiosis, including quantitative (such as intestinal bacterial overgrowth) and qualitative (such as colonic Firmicutes and Bacteroidetes levels) changes in the gut microbiota, and pathological bacterial translocation are fundamental for the pathogenesis of ALD (<a href=\"#b8-etm-0-0-09884\" rid=\"b8-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b52-etm-0-0-09884\" rid=\"b52-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">52<\/a>). A number of these intestinal microbiota are affected by alcohol, which express hydrogenases and act as the main producers of endogenous H<sub>2<\/sub> in humans and animals (<a href=\"#b10-etm-0-0-09884\" rid=\"b10-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">10<\/a>,<a href=\"#b20-etm-0-0-09884\" rid=\"b20-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>,<a href=\"#b52-etm-0-0-09884\" rid=\"b52-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">52<\/a>,<a href=\"#b53-etm-0-0-09884\" rid=\"b53-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">53<\/a>). Endogenous H<sub>2<\/sub> is crucial for hepatic redox homeostasis, glucose and lipid homeostasis, in addition to immune and inflammatory homeostasis (<a href=\"#b11-etm-0-0-09884\" rid=\"b11-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">11<\/a>,<a href=\"#b54-etm-0-0-09884\" rid=\"b54-etm-0-0-09884\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">54-57<\/a>). Therefore, it remains of importance to investigate the effects of endogenous H<sub>2<\/sub> in the pathogenesis of ALD.<\/p>\n<p class=\"p p-last\">In summary, findings of the present study indicated that intraperitoneal injection with exogenous H<sub>2<\/sub> attenuated acute alcohol-induced liver injury in mice by inhibiting hepatic JNK activation. Therefore, it would be possible to treat alcohol-induced liver injury with H<sub>2<\/sub>, such as drinking H<sub>2<\/sub>-rich alcohol or H<sub>2<\/sub>-rich water, where H<sub>2<\/sub> can be a potentially useful natural agent for the treatment of ALD.<\/p>\n<\/div>\n<div id=\"ack-a.i.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=\"ack-a.i.atitle\">Acknowledgements<\/h2>\n<div class=\"sec\">\n<p>Not applicable.<\/p>\n<\/div>\n<\/div>\n<div id=\"funding-group-a.g.b.v\" 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=\"funding-group-a.g.b.vtitle\">Funding Statement<\/h2>\n<p><strong>Funding:<\/strong> The present study was supported by National Natural Science Foundation of China (grant no. 81900376), Natural Science Foundation of Guangdong Province (grant no. 2018A030313657), the Project funded by China Postdoctoral Science Foundation (grant no. 2019M653238) and Guangdong famous Traditional Chinese Medicine inheritance studio construction project (grant no. 20180137).<\/p>\n<\/div>\n<div id=\"sec-a.i.b\" class=\"tsec bk-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=\"sec-a.i.btitle\">Availability of data and materials<\/h2>\n<p><!--\/article\/back\/sec\/--><\/p>\n<p class=\"p p-first-last\">The datasets used and\/or analyzed during the current study are available from the corresponding author on reasonable request.<\/p>\n<\/div>\n<div id=\"sec-a.i.c\" class=\"tsec bk-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=\"sec-a.i.ctitle\">Authors&#8217; contributions<\/h2>\n<p><!--\/article\/back\/sec\/--><\/p>\n<p class=\"p p-first-last\">YZ, WG, MD and HY conceived and designed the experiments. YZ, GZ, ZC, MB and YH performed the experiments and analyzed the data. YZ drafted the manuscript. All authors read and approved the final manuscript.<\/p>\n<\/div>\n<div id=\"sec-a.i.d\" class=\"tsec bk-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=\"sec-a.i.dtitle\">Ethics approval and consent to participate<\/h2>\n<p><!--\/article\/back\/sec\/--><\/p>\n<p class=\"p p-first-last\">All animal procedures were approved by the Institutional Animal Care and Use Committee of Sun Yat-sen University (IACUC code no: 2018-057).<\/p>\n<\/div>\n<div id=\"sec-a.i.e\" class=\"tsec bk-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=\"sec-a.i.etitle\">Patient consent for publication<\/h2>\n<p><!--\/article\/back\/sec\/--><\/p>\n<p class=\"p p-first-last\">Not applicable.<\/p>\n<\/div>\n<div id=\"sec-a.i.f\" class=\"tsec bk-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=\"sec-a.i.ftitle\">Competing interests<\/h2>\n<p><!--\/article\/back\/sec\/--><\/p>\n<p class=\"p p-first-last\">The authors declare that they have no competing interests.<\/p>\n<\/div>\n<div id=\"ref-list-a.i.g\" 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=\"ref-list-a.i.gtitle\">References<\/h2>\n<div class=\"ref-list-sec sec\" id=\"reference-list\">\n<div class=\"ref-cit-blk half_rhythm\" id=\"b1-etm-0-0-09884\">1. <span class=\"element-citation\">Osna NA, Donohue TM Jr, Kharbanda KK. 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href=\"https:\/\/scholar.google.com\/scholar_lookup?journal=Nutrients&amp;title=L-arabinose+elicits+gut-derived+hydrogen+production+and+ameliorates+metabolic+syndrome+in+C57BL\/6J+mice+on+high-fat-diet&amp;author=L+Zhao&amp;author=Y+Wang&amp;author=G+Zhang&amp;author=T+Zhang&amp;author=J+Lou&amp;volume=11&amp;issue=3054&amp;publication_year=2019&amp;pmid=31847305&amp;doi=10.3390\/nu11123054&amp;\" target=\"_blank\" rel=\"noopener noreferrer\" ref=\"reftype=other&amp;article-id=7976433&amp;issue-id=377760&amp;journal-id=1895&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=\"#abstract-a.g.b.ttitle\">Abstract<\/a><\/li>\n<li><a href=\"#sec-a.h.atitle\">Introduction<\/a><\/li>\n<li><a href=\"#sec-a.h.btitle\">Materials and methods<\/a><\/li>\n<li><a href=\"#sec-a.h.ctitle\">Results<\/a><\/li>\n<li><a href=\"#sec-a.h.dtitle\">Discussion<\/a><\/li>\n<li><a href=\"#ack-a.i.atitle\">Acknowledgements<\/a><\/li>\n<li><a href=\"#funding-group-a.g.b.vtitle\">Funding Statement<\/a><\/li>\n<li><a href=\"#sec-a.i.btitle\">Availability of data and materials<\/a><\/li>\n<li><a href=\"#sec-a.i.ctitle\">Authors&#8217; contributions<\/a><\/li>\n<li><a href=\"#sec-a.i.dtitle\">Ethics approval and consent to participate<\/a><\/li>\n<li><a href=\"#sec-a.i.etitle\">Patient consent for publication<\/a><\/li>\n<li><a href=\"#sec-a.i.ftitle\">Competing interests<\/a><\/li>\n<li><a href=\"#ref-list-a.i.gtitle\">References<\/a><\/li>\n<\/ul>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Hydrogen gas alleviates acute alcohol-induced liver injury by inhibiting JNK activation<\/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":[879],"body-organ":[1033],"applications":[682],"test_subjects":[1518],"report-topic":[1327],"class_list":["post-26838","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-hydrogen-health","disease-liver-disease-2","body-organ-liver-2","applications-injection-2","test_subjects-mouse-2","report-topic-alcohol-toxicity-2"],"acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v28.1 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>H2 gas prevents liver injury by inhibiting JNK activation<\/title>\n<meta name=\"description\" content=\"Hydrogen gas alleviates acute alcohol-induced liver injury by inhibiting JNK activation\" \/>\n<meta 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