{"id":27309,"date":"2024-01-03T21:45:14","date_gmt":"2024-01-03T19:45:14","guid":{"rendered":"https:\/\/hho-bulgaria.com\/h2-and-hyperoxia-combo-improves-survival-organ-damage\/"},"modified":"2024-01-29T21:12:51","modified_gmt":"2024-01-29T19:12:51","slug":"h2-and-hyperoxia-combo-improves-survival-organ-damage","status":"publish","type":"post","link":"https:\/\/hho-bulgaria.com\/en\/h2-and-hyperoxia-combo-improves-survival-organ-damage\/","title":{"rendered":"H2 and Hyperoxia Combo Improves Survival &#038; Organ Damage"},"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> 2016 Jun; 11(6): 2590\u20132596. <\/div>\n<div class=\"part2\"><span class=\"fm-vol-iss-date\">Published online 2016 Apr 6. <\/span>  <span class=\"doi\"><span>doi:&nbsp;<\/span><a href=\"\/\/doi.org\/10.3892%2Fetm.2016.3231\" target=\"_blank\" rel=\"noopener noreferrer\" ref=\"reftype=other&amp;article-id=4887775&amp;issue-id=270295&amp;journal-id=1895&amp;FROM=Article%7CFront%20Matter&amp;TO=Content%20Provider%7CCrosslink%7CDOI\">10.3892\/etm.2016.3231<\/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>PMC4887775<\/span><\/div>\n<div class=\"fm-citation-pmid\">PMID: <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/27284352\">27284352<\/a><\/div>\n<\/div>\n<\/div>\n<h1 class=\"content-title\">Combination therapy of molecular hydrogen and hyperoxia improves survival rate and organ damage in a zymosan-induced generalized inflammation model<\/h1>\n<div class=\"half_rhythm\">\n<div class=\"contrib-group fm-author\"><a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=HONG%20Y%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm140611746154128\" co-class=\"co-affbox\">YUNCHUAN HONG<\/a>,<sup>1,<\/sup><sup>*<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=SUN%20L%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm140611742934176\" co-class=\"co-affbox\">LI SUN<\/a>,<sup>2,<\/sup><sup>*<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=SUN%20R%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm140611742931792\" co-class=\"co-affbox\">RUIQIANG SUN<\/a>,<sup>3,<\/sup><sup>*<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=CHEN%20H%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm140611745883712\" co-class=\"co-affbox\">HONGGUANG CHEN<\/a>,<sup>4<\/sup> <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=YU%20Y%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm140611786136384\" co-class=\"co-affbox\">YONGHAO YU<\/a>,<sup>4<\/sup> and  <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=XIE%20K%5BAuthor%5D\" class=\"affpopup\" co-rid=\"_co_idm140611786134512\" co-class=\"co-affbox\">KELIANG XIE<\/a><sup>4,<\/sup><sup>5<\/sup><sup><\/sup><\/div>\n<div style=\"display:none\" class=\"contrib-group aff-tip\">\n<div id=\"_co_idm140611746154128\">\n<h3 class=\"no_margin\">YUNCHUAN HONG<\/h3>\n<p><sup>1<\/sup>Department of Respiratory Medicine, General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=HONG%20Y%5BAuthor%5D\">YUNCHUAN HONG<\/a><\/div>\n<\/div>\n<div id=\"_co_idm140611742934176\">\n<h3 class=\"no_margin\">LI SUN<\/h3>\n<p><sup>2<\/sup>Department of Anesthesiology, General Hospital of Beijing Military Command, Beijing 100700, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=SUN%20L%5BAuthor%5D\">LI SUN<\/a><\/div>\n<\/div>\n<div id=\"_co_idm140611742931792\">\n<h3 class=\"no_margin\">RUIQIANG SUN<\/h3>\n<p><sup>3<\/sup>Department of Anesthesiology, Tianjin Eye Hospital, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=SUN%20R%5BAuthor%5D\">RUIQIANG SUN<\/a><\/div>\n<\/div>\n<div id=\"_co_idm140611745883712\">\n<h3 class=\"no_margin\">HONGGUANG CHEN<\/h3>\n<p><sup>4<\/sup>Department of Anesthesiology, Tianjin Institute of Anesthesiology, General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=CHEN%20H%5BAuthor%5D\">HONGGUANG CHEN<\/a><\/div>\n<\/div>\n<div id=\"_co_idm140611786136384\">\n<h3 class=\"no_margin\">YONGHAO YU<\/h3>\n<p><sup>4<\/sup>Department of Anesthesiology, Tianjin Institute of Anesthesiology, General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=YU%20Y%5BAuthor%5D\">YONGHAO YU<\/a><\/div>\n<\/div>\n<div id=\"_co_idm140611786134512\">\n<h3 class=\"no_margin\">KELIANG XIE<\/h3>\n<p><sup>4<\/sup>Department of Anesthesiology, Tianjin Institute of Anesthesiology, General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China<\/p>\n<p><sup>5<\/sup>Department of Anesthesiology, Clinical Medical School of Yangzhou University, Yangzhou, Jiangsu 225001, P.R. China<\/p>\n<div>Find articles by <a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/?term=XIE%20K%5BAuthor%5D\">KELIANG XIE<\/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=\"idm140611745356640_ai\">Author information<\/a> <a href=\"#\" class=\"pmctoggle\" rid=\"idm140611745356640_an\">Article notes<\/a> <a href=\"#\" class=\"pmctoggle\" rid=\"idm140611745356640_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=\"idm140611745356640_ai\" style=\"display:none\">\n<div class=\"fm-affl\" id=\"af1-etm-0-0-3231\"><sup>1<\/sup>Department of Respiratory Medicine, General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China<\/div>\n<div class=\"fm-affl\" id=\"af2-etm-0-0-3231\"><sup>2<\/sup>Department of Anesthesiology, General Hospital of Beijing Military Command, Beijing 100700, P.R. China<\/div>\n<div class=\"fm-affl\" id=\"af3-etm-0-0-3231\"><sup>3<\/sup>Department of Anesthesiology, Tianjin Eye Hospital, Tianjin Key Lab of Ophthalmology and Visual Science, Tianjin 300020, P.R. China<\/div>\n<div class=\"fm-affl\" id=\"af4-etm-0-0-3231\"><sup>4<\/sup>Department of Anesthesiology, Tianjin Institute of Anesthesiology, General Hospital of Tianjin Medical University, Tianjin 300052, P.R. China<\/div>\n<div class=\"fm-affl\" id=\"af5-etm-0-0-3231\"><sup>5<\/sup>Department of Anesthesiology, Clinical Medical School of Yangzhou University, Yangzhou, Jiangsu 225001, P.R. China<\/div>\n<div id=\"c1-etm-0-0-3231\"><em>Correspondence to<\/em>: Dr Keliang Xie, Department of Anesthesiology, Tianjin Institute of Anesthesiology, General Hospital of Tianjin Medical University, 154 Anshan Road, Tianjin 300052, P.R. China, E-mail: <a href=\"mailto:dev@null\" data-email=\"moc.liamtoh@9002gnailekeix\" class=\"oemail\">moc.liamtoh@9002gnailekeix<\/a><\/div>\n<div id=\"fn1-etm-0-0-3231\"><sup>*<\/sup>Contributed equally<\/div>\n<\/div>\n<div class=\"fm-article-notes hide half_rhythm\" id=\"idm140611745356640_an\" style=\"display:none\">\n<div class=\"fm-pubdate half_rhythm\">Received 2014 Dec 20; Accepted 2016 Feb 11.<\/div>\n<\/div>\n<div class=\"permissions half_rhythm hide\" id=\"idm140611745356640_cpl\" style=\"display:none\">\n<div class=\"fm-copyright half_rhythm\"><a href=\"\/pmc\/about\/copyright\/\">Copyright<\/a>  \u00a9 2016, Spandidos Publications<\/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=\"abstract-a.f.b.u\" 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.f.b.utitle\">Abstract<\/h2>\n<p><!--article-meta--><\/p>\n<div>\n<p class=\"p p-first-last\">Multiple organ dysfunction syndrome (MODS) is a leading cause of mortality in critically ill patients. Hyperoxia treatment may be beneficial to critically ill patients. However, the clinical use of hyperoxia is hindered as it may exacerbate organ injury by increasing reactive oxygen species (ROS). Hydrogen gas (H<sub>2<\/sub>) exerts a therapeutic antioxidative effect by selectively reducing ROS. Combination therapy of H<sub>2<\/sub> and hyperoxia has previously been shown to significantly improve survival rate and organ damage extent in mice with polymicrobial sepsis. The aim of the present study was to investigate whether combination therapy with H<sub>2<\/sub> and hyperoxia could improve survival rate and organ damage in a zymosan (ZY)-induced generalized inflammation model. The results showed that the inhalation of H<sub>2<\/sub> (2%) or hyperoxia (98%) alone improved the 14-day survival rate of ZY-challenged mice from 20 to 70 or 60%, respectively. However, combination therapy with H<sub>2<\/sub> and hyperoxia could increase the 14-day survival rate of ZY-challenged mice to 100%. Furthermore, ZY-challenged mice showed significant multiple organ damage characterized by increased serum levels of aspartate transaminase, alanine transaminase, blood urea nitrogen and creatinine, as well as lung, liver and kidney histopathological scores at 24 h after ZY injection. These symptoms where attenuated by H<sub>2<\/sub> or hyperoxia alone; however, combination therapy with H<sub>2<\/sub> and hyperoxia had a more marked beneficial effect against lung, liver and kidney damage in ZY-challenged mice. In addition, the beneficial effects of this combination therapy on ZY-induced organ damage were associated with decreased serum levels of the oxidative product 8-iso-prostaglandin F2\u03b1, increased activity of superoxide dismutase and reduced levels of the proinflammatory cytokines high-mobility group box 1 and tumor necrosis factor-\u03b1. In conclusion, combination therapy with H<sub>2<\/sub> and hyperoxia provides enhanced therapeutic efficacy against multiple organ damage in a ZY-induced generalized inflammation model, suggesting the potential applicability of H<sub>2<\/sub> and hyperoxia in the therapy of conditions associated with inflammation-related MODS.<\/p>\n<\/div>\n<div class=\"sec\"><strong class=\"kwd-title\">Keywords: <\/strong><span class=\"kwd-text\">multiple organ dysfunction syndrome\/failure, reactive oxygen species, inflammatory cytokines, hydrogen gas, hyperoxia<\/span><\/div>\n<\/div>\n<div id=\"sec-a.g.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.g.atitle\">Introduction<\/h2>\n<p class=\"p p-first\">Multiple organ dysfunction syndrome (MODS) is the leading cause of mortality in critically ill patients (<a href=\"#b1-etm-0-0-3231\" rid=\"b1-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">1<\/a>). MODS is defined as the progressive deterioration of organ function, which often occurs in patients with severe sepsis, septic shock, hemorrhagic shock, multiple trauma, severe burns or pancreatitis (<a href=\"#b2-etm-0-0-3231\" rid=\"b2-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">2<\/a>). The mechanisms underlying the pathology of MODS are not fully understood (<a href=\"#b3-etm-0-0-3231\" rid=\"b3-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">3<\/a>), therefore it is difficult to develop an effective therapeutic measure for patients with MODS.<\/p>\n<p>Oxygen inhalation is frequently administered to critically ill patients. Early goal-directed therapy for MODS aims to balance tissue oxygen delivery and oxygen demand. Previous studies have suggested that hyperoxia may have a beneficial effect against sepsis and sepsis-associated multiple organ damage (<a href=\"#b4-etm-0-0-3231\" rid=\"b4-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">4<\/a>\u2013<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>). However, the clinical use of hyperoxia is limited in critically ill patients due to concerns that it may exacerbate organ damage by increasing free radical formation. Oxidative stress is crucially involved in the pathogenesis of MODS, and overproduction of reactive oxygen species (ROS) may exacerbate organ damage (<a href=\"#b9-etm-0-0-3231\" rid=\"b9-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">9<\/a>,<a href=\"#b10-etm-0-0-3231\" rid=\"b10-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">10<\/a>). It has been suggested that a low concentration of molecular hydrogen (H<sub>2<\/sub>) exerts a therapeutic antioxidative effect by selectively reducing hydroxyl (<sup>\u2022<\/sup>OH)radicals, the most cytotoxic ROS, and peroxynitrite (ONOO<sup>\u2212<\/sup>), and effectively protects against numerous diseases (<a href=\"#b11-etm-0-0-3231\" rid=\"b11-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">11<\/a>\u2013<a href=\"#b15-etm-0-0-3231\" rid=\"b15-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">15<\/a>). Furthermore, our previous study showed that combination therapy with H<sub>2<\/sub> and hyperoxia can significantly alleviate organ injury and improve survival rate of mice with polymicrobial sepsis via reducing oxidative stress and inflammation (<a href=\"#b16-etm-0-0-3231\" rid=\"b16-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">16<\/a>).<\/p>\n<p class=\"p p-last\">Therefore, these findings strongly indicate that combination therapy with H<sub>2<\/sub> and hyperoxia may afford a more potent therapeutic strategy for sepsis. The zymosan (ZY)-induced generalized inflammation model has been widely used by other research groups (<a href=\"#b17-etm-0-0-3231\" rid=\"b17-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">17<\/a>), as well as by our group (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>), because ZY, a substance derived from the cell wall of the yeast <em>Saccharomyces cerevisiae<\/em>, can produce systemic inflammation by inducing a wide range of inflammatory mediators (<a href=\"#b18-etm-0-0-3231\" rid=\"b18-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">18<\/a>). This model has also been used in previous experimental studies investigating MODS (<a href=\"#b18-etm-0-0-3231\" rid=\"b18-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">18<\/a>). Therefore, the aim of the present study was to investigate whether combination therapy with H<sub>2<\/sub> and hyperoxia could produce enhanced efficacy in a murine model of ZY-induced generalized inflammation.<\/p>\n<\/div>\n<div id=\"sec-a.g.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.g.btitle\">Materials and methods<\/h2>\n<div id=\"sec-a.g.b.b\" class=\"sec sec-first-last\">\n<div id=\"sec-a.g.b.b.b\" class=\"sec sec-first\">\n<h4 id=\"sec-a.g.b.b.btitle\" class=\"inline\">Animals <\/h4>\n<p class=\"p p-first-last\">Male Imprinting Control Region (ICR) mice (specific pathogen-free; age, 6\u20138 weeks; weight, 20\u201325 g) provided by the Laboratory Animal Center of the Academy of Military Science of the Chinese People&#8217;s Liberation Army (Beijing, China) were used in all experiments. Animals were housed at 20\u201322\u00b0C with a 12-h light\/dark cycle. Animals were fed standard chow and water <em>ad libitum<\/em>. All experimental protocols were approved by the Institutional Animal Care and Use Committee of Tianjin Medical University (Tianjin, China), and were performed in accordance with the National Institutes of Health guidelines for the use of experimental animals (<a href=\"#b19-etm-0-0-3231\" rid=\"b19-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">19<\/a>).<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.c\" class=\"sec\">\n<h4 id=\"sec-a.g.b.b.ctitle\" class=\"inline\">ZY-induced generalized inflammation model <\/h4>\n<p class=\"p p-first-last\">ZY (Sigma-Aldrich, St. Louis, MO, USA) was dissolved in an isotonic sodium chloride solution (normal saline [NS]) to a final concentration of 25 mg\/ml and was sterilized at 100\u00b0C for 80 min. All suspensions were freshly produced prior to use. Generalized inflammation was induced by an aseptic intraperitoneal injection of ZY (1 g\/kg) (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b18-etm-0-0-3231\" rid=\"b18-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">18<\/a>). The same volume of NS was injected via the same route as a control.<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.d\" class=\"sec\">\n<h4 id=\"sec-a.g.b.b.dtitle\" class=\"inline\">Molecular hydrogen and\/or hyperoxia treatment <\/h4>\n<p class=\"p p-first-last\">The animals were put in a sealed plexiglass chamber with inflow and outflow outlets (<a href=\"#b20-etm-0-0-3231\" rid=\"b20-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>,<a href=\"#b21-etm-0-0-3231\" rid=\"b21-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">21<\/a>). H<sub>2<\/sub>, O<sub>2<\/sub> or N<sub>2<\/sub> was supplied via a gas flowmeter, (Yukata Engineering Corp., Tokyo, Japan) and delivered into the chamber through a tube at a rate of 4 l\/min. The concentrations of O<sub>2<\/sub> and H<sub>2<\/sub> in the chamber were continuously monitored using a gas analyzer (LB-2, Model 40 M; Beckman Coulter, Inc., Fullerton, CA, USA) and a commercially available hydrogen detector (HY-ALERTA\u2122 500, H2scan Corporation, Valencia, CA, USA), respectively. CO<sub>2<\/sub> was removed from the chamber gases using baralyme (Allied Healthcare Products, Inc., St. Louis, MO, USA). The mixed gases were maintained at the following levels: 2% H<sub>2<\/sub>, 21% O<sub>2<\/sub> and 77% N<sub>2<\/sub&gt;; 0% H<sub>2<\/sub>, 98% O<sub>2<\/sub> and 2% N<sub>2<\/sub&gt;; and 2% H<sub>2<\/sub>, 98% O<sub>2<\/sub> and 0% N<sub>2<\/sub> during the treatment. The animals without H<sub>2<\/sub> or hyperoxia treatment were exposed to room air in the chamber.<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.e\" class=\"sec sec-last\">\n<h4 id=\"sec-a.g.b.b.etitle\" class=\"inline\">Experimental design <\/h4>\n<div id=\"sec-a.g.b.b.e.b\" class=\"sec sec-first\">\n<h5 id=\"sec-a.g.b.b.e.btitle\" class=\"inline\">Experiment one: Effects of H<sub>2<\/sub> and\/or hyperoxia treatment on the survival rate in ZY-challenged mice <\/h5>\n<p class=\"p p-first-last\">A total of 180 animals were randomly allocated into six groups (n=30 per group): Normal saline (NS), NS + H<sub>2<\/sub> + O<sub>2<\/sub&gt;; ZY; ZY + H<sub>2<\/sub&gt;; ZY + O<sub>2<\/sub&gt;; and ZY + H<sub>2<\/sub> + O<sub>2<\/sub> groups. The treatment concentrations of hydrogen and hyperoxia were determined based on our previous studies and preliminary observations (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b21-etm-0-0-3231\" rid=\"b21-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">21<\/a>). The animals in the ZY + O<sub>2<\/sub> and ZY + H<sub>2<\/sub> groups were exposed to 98% O<sub>2<\/sub> or 2% H<sub>2<\/sub> for 3 h starting at 1 and 6 h after ZY injection, respectively. The animals in the NS + H<sub>2<\/sub> + O<sub>2<\/sub> and ZY + H<sub>2<\/sub> + O<sub>2<\/sub> groups were exposed to 98% O<sub>2<\/sub> and 2% H<sub>2<\/sub> at the same time points. As a control, the animals from the NS and ZY groups were given room air treatment at the same time points. The survival rate was observed on days 1, 2, 3, 4, 5, 6, 7 and 14 after NS or ZY injection. In addition, arterial blood gas evaluation was conducted at 1.5 h after the onset of H<sub>2<\/sub> inhalation (2.5 h after NS or ZY injection) in all groups.<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.e.c\" class=\"sec\">\n<h5 id=\"sec-a.g.b.b.e.ctitle\" class=\"inline\">Experiment two: Effects of H<sub>2<\/sub> and\/or hyperoxia treatment on organ injury in ZY-challenged mice <\/h5>\n<p class=\"p p-first\">Additional 36 animals were used in this experiment and were assigned to six groups (n=6 per group). The grouping method and experimental protocols were the same as Experiment One. At 24 h after NS or ZY injection, all the animals were sacrificed with sodium pentobarbital (50 mg\/kg, intraperitoneally), and the blood samples and organs were collected for detecting serum biochemical parameters and organ histopathology.<\/p>\n<p class=\"p p-last\">In addition, at 24 h after NS or ZY injection, the serum levels of the proinflammatory cytokines high-mobility group box 1 (HMGB1) and tumor necrosis factor-\u03b1 (TNF-\u03b1), the antioxidative enzyme superoxide dismutase (SOD) and the oxidative product 8-iso-prostaglandin F2\u03b1 (8-iso-PGF2\u03b1) were evaluated.<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.e.d\" class=\"sec\">\n<h5 id=\"sec-a.g.b.b.e.dtitle\" class=\"inline\">Arterial blood gas analysis <\/h5>\n<p class=\"p p-first-last\">The arterial blood gas analysis (for pH, PaO<sub>2<\/sub> and PaCO<sub>2<\/sub>) was conducted using a GEM Premier 3000 gas analyzer (Instrumentation Laboratory Spa, Milan, Italy).<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.e.e\" class=\"sec\">\n<h5 id=\"sec-a.g.b.b.e.etitle\" class=\"inline\">Serum biochemical parameters assay <\/h5>\n<p class=\"p p-first-last\">The serum was separated from whole blood samples by centrifugation at 1,100 \u00d7 g for 10 min, aliquoted and stored at \u221280\u00b0C until assayed (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b20-etm-0-0-3231\" rid=\"b20-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>\u2013<a href=\"#b22-etm-0-0-3231\" rid=\"b22-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>). The samples were evaluated using a biochemistry autoanalyzer (7150; Hitachi, Ltd., Tokyo, Japan) to measure serum levels of alanine aminotransferase (ALT, IU\/l), aspartate aminotransferase (AST, IU\/l), blood urea nitrogen (BUN, mmol\/l) and creatinine (Cr, \u00b5mol\/l).<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.e.f\" class=\"sec\">\n<h5 id=\"sec-a.g.b.b.e.ftitle\" class=\"inline\">Organ histological examination <\/h5>\n<p class=\"p p-first-last\">The lung, liver and kidneys were removed immediately, fixed in 4% paraformaldehyde (Sigma-Aldrich), embedded in paraffin, and sectioned at 4\u20136 \u00b5m. After deparaffinization and rehydration, the sections were stained with hematoxylin and eosin (Sigma-Aldrich). Based on the scoring standard in our previous studies (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b22-etm-0-0-3231\" rid=\"b22-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>), the histological slides were blindly examined and scored by two experienced pathologists.<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.e.g\" class=\"sec\">\n<h5 id=\"sec-a.g.b.b.e.gtitle\" class=\"inline\">Detection of SOD activity <\/h5>\n<p class=\"p p-first-last\">The activities of SOD were measured using commercial kits purchased from Cayman Chemical Company (Ann Arbor, MI, USA). Total SOD activity was assayed according to the manufacturer&#8217;s instructions and the protocol described in our previous studies (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b20-etm-0-0-3231\" rid=\"b20-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>\u2013<a href=\"#b22-etm-0-0-3231\" rid=\"b22-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>). All spectrophotometric readings were performed using a spectrophotometer (DU-640B; Beckman Coulter, Inc.). All assays were conducted in triplicates.<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.e.h\" class=\"sec\">\n<h5 id=\"sec-a.g.b.b.e.htitle\" class=\"inline\">Detection of 8-iso-PGF2\u03b1 <\/h5>\n<p class=\"p p-first-last\">Measurement of 8-iso-PGF2\u03b1, a free radical-catalyzed product of arachidonic acid, can offer a reliable approach for quantitative measurement of oxidative stress status <em>in vivo<\/em> (<a href=\"#b23-etm-0-0-3231\" rid=\"b23-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">23<\/a>). The levels of serum 8-iso-PGF2\u03b1 were detected using specific enzyme-linked immunosorbent assay (ELISA) kits (cat. no. 516351; Cayman Chemical Company) and a Spectramax M5 microplate reader (Molecular Devices, LLC, Sunnyvale, CA, USA) (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b20-etm-0-0-3231\" rid=\"b20-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>\u2013<a href=\"#b22-etm-0-0-3231\" rid=\"b22-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>). All standards and samples were run in duplicate.<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.e.i\" class=\"sec\">\n<h5 id=\"sec-a.g.b.b.e.ititle\" class=\"inline\">Detection of inflammatory cytokines <\/h5>\n<p class=\"p p-first-last\">The levels of serum TNF-\u03b1 (R&amp;D Systems, Inc., Minneapolis, MI, USA) and HMGB1 (IBL International GmbH, Hamburg, Germany) were detected using specific ELISA kits and the Molecular Devices microplate reader (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b20-etm-0-0-3231\" rid=\"b20-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>\u2013<a href=\"#b22-etm-0-0-3231\" rid=\"b22-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>). All standards and samples were run in duplicate.<\/p>\n<\/div>\n<div id=\"sec-a.g.b.b.e.j\" class=\"sec sec-last\">\n<h5 id=\"sec-a.g.b.b.e.jtitle\" class=\"inline\">Statistical analysis <\/h5>\n<p class=\"p p-first-last\">Survival rates are expressed as percentages and measurement data are expressed as the mean \u00b1 standard error of the mean. The analysis of survival rates were tested using Fisher&#8217;s exact probability method. The inter-group differences of the rest data were tested by one-way analysis of variance followed by Least Significant Difference t-test for multiple comparisons. Statistical analyses were performed using SPSS software, version 16.0 (SPSS, Inc., Chicago, IL, USA). P&lt;0.05 was considered to indicate a statistically significant difference.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.g.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.g.ctitle\">Results<\/h2>\n<div id=\"sec-a.g.c.b\" class=\"sec sec-first-last\">\n<div id=\"sec-a.g.c.b.b\" class=\"sec sec-first\">\n<h4 id=\"sec-a.g.c.b.btitle\" class=\"inline\">Differences in arterial blood gas during treatment <\/h4>\n<p class=\"p p-first-last\">In the present study, the effects of H<sub>2<\/sub> inhalation on arterial pH, PaO<sub>2<\/sub> and PaCO<sub>2<\/sub> were evaluated in all groups at 1.5 h after the onset of H<sub>2<\/sub> and\/or O<sub>2<\/sub> inhalation (2.5 h after NS or ZY injection). The levels of P<sub>a<\/sub>O<sub>2<\/sub> in the ZY + O<sub>2<\/sub>, ZY + H<sub>2<\/sub> + O<sub>2<\/sub> and NS + H<sub>2<\/sub> + O<sub>2<\/sub> groups were 405.1\u00b119.2, 410.3\u00b118.7 and 408.4\u00b121.5 mmHg, respectively. The P<sub>a<\/sub>O<sub>2<\/sub> levels in the NS, ZY and ZY + H<sub>2<\/sub> groups were 97.18\u00b18.29, 96.72\u00b110.31 and 95.46\u00b18.46 mmHg, respectively. There were no differences in the levels of arterial pH and P<sub>a<\/sub>CO<sub>2<\/sub> among all groups (<em>data not shown<\/em>). The results demonstrate that hyperoxia exposure can significantly increase the P<sub>a<\/sub>O<sub>2<\/sub> level, and 2% H<sub>2<\/sub> has no marked effects on the P<sub>a<\/sub>O<sub>2<\/sub> level in ZY-challenged mice during the treatment.<\/p>\n<\/div>\n<div id=\"sec-a.g.c.b.c\" class=\"sec\">\n<h4 id=\"sec-a.g.c.b.ctitle\" class=\"inline\">Combination therapy with H<sub>2<\/sub> and hyperoxia improves the survival rate in ZY-challenged mice <\/h4>\n<p class=\"p p-first-last\">The 14-day survival rate of ZY-challenged mice was 20% (P&lt;0.05 vs. NS group; <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f1-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f1-etm-0-0-3231\" rid-ob=\"ob-f1-etm-0-0-3231\" co-legend-rid=\"lgnd_f1-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 1<\/span><\/a>). The results showed that either 98% O<sub>2<\/sub> or 2% H<sub>2<\/sub> exposure for 3 h starting at 1 and 6 h after ZY injection, respectively, improved the 14-day survival rate of ZY-challenged mice to 60 or 70% (P&lt;0.05 vs. ZY group; <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f1-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f1-etm-0-0-3231\" rid-ob=\"ob-f1-etm-0-0-3231\" co-legend-rid=\"lgnd_f1-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 1<\/span><\/a>). Furthermore, combination therapy with 2% H<sub>2<\/sub> and 98% O<sub>2<\/sub> increased the 14-day survival rate of ZY-challenged mice to 100% (P&lt;0.05 vs. ZY group; <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f1-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f1-etm-0-0-3231\" rid-ob=\"ob-f1-etm-0-0-3231\" co-legend-rid=\"lgnd_f1-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 1<\/span><\/a>). In addition, all mice in the NS and NS + H<sub>2<\/sub> + O<sub>2<\/sub> groups survived during the experiment. These results suggest that combination therapy with H<sub>2<\/sub> and hyperoxia can improve the survival rate of ZY-challenged mice in a synergistic manner.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"f1-etm-0-0-3231\" co-legend-rid=\"lgnd_f1-etm-0-0-3231\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f1-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f1-etm-0-0-3231\" rid-ob=\"ob-f1-etm-0-0-3231\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm140611744905648\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f1-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f1-etm-0-0-3231\" rid-ob=\"ob-f1-etm-0-0-3231\" 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=4887775_etm-11-06-2590-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-11-06-2590-g00.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/etm-11-06-2590-g00.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm140611744905648\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f1-etm-0-0-3231\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_f1-etm-0-0-3231\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f1-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f1-etm-0-0-3231\" rid-ob=\"ob-f1-etm-0-0-3231\" rel=\"noopener\">Figure 1.<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of H<sub>2<\/sub> and\/or hyperoxia on the survival rate of zymosan-challenged mice. The mice were treated with or without 2% H<sub>2<\/sub> and\/or 98% O<sub>2<\/sub> inhalation for 3 h starting at 1 and 6 h after normal saline or zymosan injection, respectively. Values are expressed as survival percentage (n=30 per group). *P&lt;0.05 vs. ZY group; <sup>\u2020<\/sup>P&lt;0.05 vs. ZY + H<sub>2<\/sub> group; <sup>\u2021<\/sup>P&lt;0.05 vs. ZY + O<sub>2<\/sub> group. ZY, zymosan; d, days.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.g.c.b.d\" class=\"sec\">\n<h4 id=\"sec-a.g.c.b.dtitle\" class=\"inline\">Combination therapy with H<sub>2<\/sub> and hyperoxia improves organ histopathological scores in ZY-challenged mice <\/h4>\n<p class=\"p p-first-last\">At 24 h after ZY or NS injection, the animals in all groups were killed for histopathological analysis. According to the scoring standard in our previous studies (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b22-etm-0-0-3231\" rid=\"b22-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>), the histopathological changes in lung, liver and kidney were scored using a scale of 1 to 4. As shown in the <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f2-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f2-etm-0-0-3231\" rid-ob=\"ob-f2-etm-0-0-3231\" co-legend-rid=\"lgnd_f2-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 2<\/span><\/a>, the histopathological scores for lung, liver and kidney in the ZY group were 3\u20133.5, significantly increased compared with the NS group (P&lt;0.05). These abnormal changes in ZY-challenged mice were attenuated by either 98% O<sub>2<\/sub> or 2% H<sub>2<\/sub> treatment alone (P&lt;0.05 vs. ZY group; <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f2-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f2-etm-0-0-3231\" rid-ob=\"ob-f2-etm-0-0-3231\" co-legend-rid=\"lgnd_f2-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 2<\/span><\/a>). Furthermore, these abnormal changes in ZY-challenged mice were more markedly ameliorated by combination therapy with 98% O<sub>2<\/sub> and 2% H<sub>2<\/sub> compared with either treatment alone (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f2-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f2-etm-0-0-3231\" rid-ob=\"ob-f2-etm-0-0-3231\" co-legend-rid=\"lgnd_f2-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 2<\/span><\/a>). These data indicate that combination therapy with H<sub>2<\/sub> and hyperoxia has an enhanced efficacy against multiple organ damage in ZY-challenged mice.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"f2-etm-0-0-3231\" co-legend-rid=\"lgnd_f2-etm-0-0-3231\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f2-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f2-etm-0-0-3231\" rid-ob=\"ob-f2-etm-0-0-3231\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm140611744922544\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f2-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f2-etm-0-0-3231\" rid-ob=\"ob-f2-etm-0-0-3231\" 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=4887775_etm-11-06-2590-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-11-06-2590-g01.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/etm-11-06-2590-g01.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm140611744922544\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f2-etm-0-0-3231\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_f2-etm-0-0-3231\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f2-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f2-etm-0-0-3231\" rid-ob=\"ob-f2-etm-0-0-3231\" rel=\"noopener\">Figure 2.<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of H<sub>2<\/sub> and\/or hyperoxia on the organ histopathological scores in zymosan-challenged mice. Histological scores in the mouse (A) lungs, (B) liver and (C) kidneys. Mice were treated with or without 2% H<sub>2<\/sub> and\/or 98% O<sub>2<\/sub> inhalation for 3 h starting at 1 and 6 h after NS or zymosan injection, respectively. At 24 h after NS or zymosan injection, all animals were anesthetized, and the organ samples were collected for measuring the histopathological scores. Values are expressed as the mean \u00b1 standard error of the mean (n=6 per group). *P&lt;0.05 vs. NS group; <sup>\u2020<\/sup>P&lt;0.05 vs. ZY group; <sup>\u2021<\/sup>P&lt;0.05 vs. ZY + O<sub>2<\/sub> group. NS, normal saline; ZY, zymosan.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.g.c.b.e\" class=\"sec\">\n<h4 id=\"sec-a.g.c.b.etitle\" class=\"inline\">Combination therapy with H<sub>2<\/sub> and hyperoxia improves serum biochemical parameters in ZY-challenged mice <\/h4>\n<p class=\"p p-first-last\">As shown in <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f3-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f3-etm-0-0-3231\" rid-ob=\"ob-f3-etm-0-0-3231\" co-legend-rid=\"lgnd_f3-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 3<\/span><\/a>, the ZY-challenged mice appeared significantly impaired liver and kidney function at 24 h, which was assessed by serum biochemical parameters for liver and kidney function (ALT, AST, Cr and BUN). The ZY-challenged mice showed a significant increase in the levels of serum ALT, AST, Cr and BUN (P&lt;0.05 vs. NS group), which were significantly attenuated by 2% H<sub>2<\/sub> or 98% O<sub>2<\/sub> treatment alone (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f3-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f3-etm-0-0-3231\" rid-ob=\"ob-f3-etm-0-0-3231\" co-legend-rid=\"lgnd_f3-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 3<\/span><\/a>). Furthermore, these abnormal changes of biochemical parameters in ZY-challenged mice were more notably ameliorated by combination therapy with 98% O<sub>2<\/sub> and 2% H<sub>2<\/sub> (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f3-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f3-etm-0-0-3231\" rid-ob=\"ob-f3-etm-0-0-3231\" co-legend-rid=\"lgnd_f3-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 3<\/span><\/a>). These results demonstrate that combination therapy with H<sub>2<\/sub> and hyperoxia has a more substantially beneficial effect on liver and kidney dysfunction in ZY-challenged mice.<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"f3-etm-0-0-3231\" co-legend-rid=\"lgnd_f3-etm-0-0-3231\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f3-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f3-etm-0-0-3231\" rid-ob=\"ob-f3-etm-0-0-3231\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm140611750773680\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f3-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f3-etm-0-0-3231\" rid-ob=\"ob-f3-etm-0-0-3231\" 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=4887775_etm-11-06-2590-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-11-06-2590-g02.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/etm-11-06-2590-g02.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm140611750773680\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f3-etm-0-0-3231\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_f3-etm-0-0-3231\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f3-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f3-etm-0-0-3231\" rid-ob=\"ob-f3-etm-0-0-3231\" rel=\"noopener\">Figure 3.<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of H<sub>2<\/sub> and\/or hyperoxia on the serum biochemical parameters in zymosan-challenged mice. Serum levels of (A) ALT, (B) AST, (C) Cr and (D) BUN. Mice were treated with or without 2% H<sub>2<\/sub> and\/or 98% O<sub>2<\/sub> inhalation for 3 h starting at 1 and 6 h after NS or zymosan injection, respectively. At 24 h after NS or zymosan injection, all the animals were anesthetized, and the blood samples were collected for detection of the serum biochemical parameters. Values represent the mean \u00b1 standard error of the mean (n=6 per group). *P&lt;0.05 vs. NS group; <sup>\u2020<\/sup>P&lt;0.05 vs. ZY group; <sup>\u2021<\/sup>P&lt;0.05 vs. ZY + H<sub>2<\/sub> group; <sup>\u00a7<\/sup>P&lt;0.05 vs. ZY + O<sub>2<\/sub> group. ALT, alanine aminotransferase; AST, aspartate aminotransferase; BUN, blood urea nitrogen; Cr, creatinine; UI\/L, international units per liter; NS, normal saline; ZY, zymosan.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.g.c.b.f\" class=\"sec sec-last\">\n<h4 id=\"sec-a.g.c.b.ftitle\" class=\"inline\">Combination therapy with H<sub>2<\/sub> and hyperoxia prevents the abnormal changes of antioxidant enzymatic activities, oxidative product and inflammatory cytokines in ZY-challenged mice <\/h4>\n<p class=\"p p-first\">The activity of the antioxidative enzyme SOD, the levels of the oxidative product 8-iso-PGF2\u03b1 and the levels of proinflammatory cytokines (HMGB1 and TNF-\u03b1) in serum of all animals were observed at 24 h after NS or ZY injection. The results showed decreased SOD activity and increased levels of 8-iso-PGF2\u03b1, HMGB1 and TNF-\u03b1 in the serum of the ZY-challenged mice (P&lt;0.05 vs. NS group; <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f4-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f4-etm-0-0-3231\" rid-ob=\"ob-f4-etm-0-0-3231\" co-legend-rid=\"lgnd_f4-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 4<\/span><\/a>). Furthermore, the results showed that 2% H<sub>2<\/sub> or 98% O<sub>2<\/sub> alone significantly increased the SOD activity and decreased the 8-iso-PGF2\u03b1 level in serum of ZY-challenged mice (P&lt;0.05; <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f4-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f4-etm-0-0-3231\" rid-ob=\"ob-f4-etm-0-0-3231\" co-legend-rid=\"lgnd_f4-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 4<\/span><\/a>). In addition, 2% H<sub>2<\/sub> or 98% O<sub>2<\/sub> alone significantly decreased the serum HMGB1 and TNF-\u03b1 levels of the ZY-challenged mice (P&lt;0.05; <a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f4-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f4-etm-0-0-3231\" rid-ob=\"ob-f4-etm-0-0-3231\" co-legend-rid=\"lgnd_f4-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 4<\/span><\/a>). These abnormal changes in oxidative stress and inflammatory cytokines were more markedly improved by combination therapy with 98% O<sub>2<\/sub> and 2% H<sub>2<\/sub> (<a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f4-etm-0-0-3231\/\" target=\"figure\" class=\"fig-table-link figpopup\" rid-figpopup=\"f4-etm-0-0-3231\" rid-ob=\"ob-f4-etm-0-0-3231\" co-legend-rid=\"lgnd_f4-etm-0-0-3231\" rel=\"noopener\"><span>Fig. 4<\/span><\/a>).<\/p>\n<p><!--fig ft0--><!--fig mode=article f1--><\/p>\n<div class=\"fig iconblock whole_rhythm\" id=\"f4-etm-0-0-3231\" co-legend-rid=\"lgnd_f4-etm-0-0-3231\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f4-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f4-etm-0-0-3231\" rid-ob=\"ob-f4-etm-0-0-3231\" rel=\"noopener\"><!--fig\/graphic|fig\/alternatives\/graphic mode=\"anchored\" m1--><\/a><\/p>\n<div class=\"figure\" data-largeobj=\"\" data-largeobj-link-rid=\"largeobj_idm140611748728128\"><a href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f4-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f4-etm-0-0-3231\" rid-ob=\"ob-f4-etm-0-0-3231\" 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=4887775_etm-11-06-2590-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-11-06-2590-g03.jpg\" title=\"Click on image to zoom\" class=\"tileshop\" src=\"https:\/\/hho-bulgaria.com\/wp-content\/uploads\/2024\/01\/etm-11-06-2590-g03.jpg\"><\/a><\/div>\n<div class=\"largeobj-link align_right\" id=\"largeobj_idm140611748728128\" style=\"display: none;\"><a target=\"object\" rel=\"noopener\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f4-etm-0-0-3231\/?report=objectonly\">Open in a separate window<\/a><\/div>\n<div class=\"icnblk_cntnt\" id=\"lgnd_f4-etm-0-0-3231\">\n<div><a class=\"figpopup\" href=\"https:\/\/www.ncbi.nlm.nih.gov\/pmc\/articles\/PMC4887775\/figure\/f4-etm-0-0-3231\/\" target=\"figure\" rid-figpopup=\"f4-etm-0-0-3231\" rid-ob=\"ob-f4-etm-0-0-3231\" rel=\"noopener\">Figure 4.<\/a><\/div>\n<p><!--caption a7--><\/p>\n<div class=\"caption\">\n<p>Effects of H<sub>2<\/sub> and\/or hyperoxia on the activity of serum antioxidant enzyme as well as the levels of serum oxidative product and inflammatory cytokines in zymosan-challenged mice. (A) SOD activity and serum levels of (B) 8-iso-PGF2\u03b1, (C) TNF-\u03b1 and (D) HMGB1 level. The mice were treated with or without 2% H<sub>2<\/sub> and\/or 98% O<sub>2<\/sub> inhalation for 3 h starting at 1 and 6 h after NS or zymosan injection, respectively. The serum was harvested for measuring these indicators at 24 h after NS or zymosan injection. Values are expressed as the mean \u00b1 standard error of the mean (n=6 per group). *P&lt;0.05 vs. NS group; <sup>\u2020<\/sup>P&lt;0.05 vs. ZY group; <sup>\u2021<\/sup>P&lt;0.05 vs. ZY + H<sub>2<\/sub> group; <sup>\u00a7<\/sup>P&lt;0.05 vs. ZY + O<sub>2<\/sub> group. SOD, superoxide dismutase; 8-iso-PGF2\u03b1, 8-iso-prostaglandin F2\u03b1; HMGB1, high-mobility group box 1 protein; NS, normal saline; ZY, zymosan.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<p class=\"p p-last\">These data suggest that combination therapy with H<sub>2<\/sub> and hyperoxia produces a more beneficial effect on survival rate and organ damage in ZY-challenged mice, which is associated with decreased levels of oxidative product and proinflammatory cytokines and increased levels of antioxidative enzymes in the serum.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"sec-a.g.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.g.dtitle\">Discussion<\/h2>\n<p class=\"p p-first\">The present study demonstrated that either H<sub>2<\/sub> or hyperoxia treatment alone improved the survival rate of ZY-challenged mice, while combination therapy with H<sub>2<\/sub> and hyperoxia could synergistically increase the survival rate of ZY-challenged mice, which was greater than treatment with either gas alone. Furthermore, H<sub>2<\/sub> or hyperoxia treatment alone protected against lung, liver and kidney damage in ZY-challenged mice, while combination therapy with H<sub>2<\/sub> and hyperoxia provided cumulative protection against these organ damage of ZY-challenged mice. In addition, the beneficial effects of combination therapy with H<sub>2<\/sub> and hyperoxia on ZY-induced organ injury were associated with decreased levels of oxidative product 8-iso-PGF2\u03b1, increased SOD activity and reduced levels of inflammatory cytokines TNF-\u03b1 and HMGB1 in serum.<\/p>\n<p>ZY is a substance derived from the cell wall of the yeast <em>Saccharomyces cerevisiae<\/em>, and can induce systemic inflammation by inducing a wide range of inflammatory mediators (<a href=\"#b18-etm-0-0-3231\" rid=\"b18-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">18<\/a>). Based on previous studies by the present authors and others, intraperitoneal injection of a high dose of ZY (0.8\u20131.0 g\/kg) can induce a generalized inflammation in rats or mice, which is accompanied by multiple organ damage (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b17-etm-0-0-3231\" rid=\"b17-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">17<\/a>,<a href=\"#b18-etm-0-0-3231\" rid=\"b18-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">18<\/a>,<a href=\"#b22-etm-0-0-3231\" rid=\"b22-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>). ZY (1.0 g\/kg, intraperitoneal injection) successfully induced sterile inflammation model in mice, characterized by the decrease of survival rates, histopathological injury, organ dysfunction and abnormally decreased tissue oxygenation (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b22-etm-0-0-3231\" rid=\"b22-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>). In the present study, similar changes were observed in the ZY-challenged mice.<\/p>\n<p>Oxygen therapy is widely used in clinical practice as a mainstay of supportive treatment for patients with hypoxemia and critical illness. It is well known that early goal-directed therapy for MODS aims to balance oxygen delivery and demand (<a href=\"#b24-etm-0-0-3231\" rid=\"b24-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">24<\/a>,<a href=\"#b25-etm-0-0-3231\" rid=\"b25-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">25<\/a>). Previous animal studies have shown that hyperoxia exposure can improve organ function and survival rate in several models of shock or sepsis (<a href=\"#b4-etm-0-0-3231\" rid=\"b4-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">4<\/a>\u2013<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b26-etm-0-0-3231\" rid=\"b26-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">26<\/a>,<a href=\"#b27-etm-0-0-3231\" rid=\"b27-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">27<\/a>). In addition, 100% oxygen exposure for 2 and 3 h starting at 4 and 12 h, respectively, after ZY injection benefits the outcome of mice with sterile sepsis (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>). It is speculated that improved tissue oxygenation and decreased systemic inflammatory response are crucially involved in the protective effects of hyperoxia treatment (<a href=\"#b4-etm-0-0-3231\" rid=\"b4-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">4<\/a>\u2013<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>). However, hyperoxia treatment can induce the production of ROS, which are considered to be associated with oxygen toxicity (<a href=\"#b9-etm-0-0-3231\" rid=\"b9-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">9<\/a>,<a href=\"#b10-etm-0-0-3231\" rid=\"b10-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">10<\/a>). Therefore, the use of hyperoxia is limited in critically ill patients due to concerns that it may exacerbate organ damage by increasing free radical formation.<\/p>\n<p>H<sub>2<\/sub> has been used medically to prevent decompression sickness in deep divers for safety profiles (<a href=\"#b28-etm-0-0-3231\" rid=\"b28-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">28<\/a>). Prior studies have shown that H<sub>2<\/sub> exerts a therapeutic antioxidative effect by selectively reducing ROS toxicity, and effectively protecting against a number of diseases, suggesting that H<sub>2<\/sub> has potential as an antioxidant for therapeutic applications (<a href=\"#b11-etm-0-0-3231\" rid=\"b11-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">11<\/a>\u2013<a href=\"#b15-etm-0-0-3231\" rid=\"b15-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">15<\/a>). Our previous studies have shown that H<sub>2<\/sub> treatment has a beneficial effect on sepsis and sepsis-induced organ injury (<a href=\"#b16-etm-0-0-3231\" rid=\"b16-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">16<\/a>,<a href=\"#b20-etm-0-0-3231\" rid=\"b20-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>,<a href=\"#b21-etm-0-0-3231\" rid=\"b21-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">21<\/a>). In the present study, combination therapy with H<sub>2<\/sub> and hyperoxia appeared to produce a synergistic protective effect against sepsis and sepsis-associated multiple organ damage (<a href=\"#b16-etm-0-0-3231\" rid=\"b16-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">16<\/a>). The present results showed that combination therapy with H<sub>2<\/sub> and hyperoxia had a more beneficial effect on multiple organ dysfunction\/failure in the ZY-induced generalized inflammation model.<\/p>\n<p>To further investigate the possible underlying mechanisms, the effects of H<sub>2<\/sub> and\/or hyperoxia treatment on oxidant and antioxidant system in ZY-challenged mice were investigated. In our previous studies, we found that the activities of SOD, CAT and GSH-Px in serum and tissues are significantly decreased during the early and late stages of ZY-induced organ damage, indicating that ZY sets up an environment favorable for oxidative stress (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b22-etm-0-0-3231\" rid=\"b22-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>). In the present study, decreased SOD activity and increased 8-iso-PGF2\u03b1 levels were detected in the serum at 24 h after ZY injection. Furthermore, 2% H<sub>2<\/sub> and\/or 98% O<sub>2<\/sub> treatment significantly improved SOD activity and decreased the 8-iso-PGF2\u03b1 levels in the serum. These results suggest that the reduction of oxidative damage and the increase of endogenous antioxidant enzymatic activities in the serum may associated with the protective effects of H<sub>2<\/sub> and\/or O<sub>2<\/sub> treatment, which is consistent with our previous study (<a href=\"#b20-etm-0-0-3231\" rid=\"b20-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>). It has been hypothesized that the uncontrolled and exaggerated inflammatory response plays a major role in the pathogenesis of sepsis\/MODS (<a href=\"#b3-etm-0-0-3231\" rid=\"b3-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">3<\/a>). Inflammatory cytokines include early inflammatory cytokines such as proinflammatory cytokines TNF-\u03b1, IL-6 and anti-inflammatory cytokine IL-10, as well as the late inflammatory cytokine HMGB1 (<a href=\"#b29-etm-0-0-3231\" rid=\"b29-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">29<\/a>,<a href=\"#b30-etm-0-0-3231\" rid=\"b30-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">30<\/a>). Early and late inflammatory cytokines can interact and facilitate the organ dysfunction and injury in sepsis\/MODS. Our previous studies have demonstrated that HMGB1 contributed to organ damage in the ZY-induced generalized inflammation model (<a href=\"#b8-etm-0-0-3231\" rid=\"b8-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">8<\/a>,<a href=\"#b22-etm-0-0-3231\" rid=\"b22-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">22<\/a>). In the present study, we found that ZY-challenged mice showed the significant increase of TNF-\u03b1 and HMGB1 in serum, which was significantly attenuated by 2% H<sub>2<\/sub> and\/or O<sub>2<\/sub> treatment. These data suggest that the protective effects of combination therapy with H<sub>2<\/sub> and hyperoxia treatment on ZY-challenged mice are associated with a reduction in the serum levels of early and late proinflammatory cytokines, which is consistent with our previous study (<a href=\"#b20-etm-0-0-3231\" rid=\"b20-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">20<\/a>,<a href=\"#b21-etm-0-0-3231\" rid=\"b21-etm-0-0-3231\" class=\" bibr popnode\" role=\"button\" aria-expanded=\"false\" aria-haspopup=\"true\">21<\/a>).<\/p>\n<p class=\"p p-last\">H<sub>2<\/sub> is neither explosive nor dangerous at a low concentration (&lt;4.1% in oxygen). The present results suggest that a mixed gas therapy with H<sub>2<\/sub> and hyperoxia may be a novel, safe and effective approach for preventing organ damage and mortality in MODS.<\/p>\n<\/div>\n<div id=\"ack-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=\"ack-a.h.atitle\">Acknowledgements<\/h2>\n<div class=\"sec\">\n<p>This study was supported by research grants from the National Natural Science Foundation of China (grant nos. 81101409 and 81471842) and Natural Science Foundation of Tianjin, China (no. 13JCQNJC11400), the Foundation of Tianjin Bureau of Public Health (no. 2011KZ108).<\/p>\n<\/div>\n<\/div>\n<div id=\"glossary-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=\"glossary-a.h.btitle\">Glossary<\/h2>\n<div class=\"bk-sec\">\n<div>\n<h4>Abbreviations<\/h4>\n<table class=\"default-table glossary\">\n<tbody>\n<tr>\n<td>8-iso-PGF2\u03b1<\/td>\n<td>8-iso-prostaglandin F2\u03b1<\/td>\n<\/tr>\n<tr>\n<td>ALT<\/td>\n<td>alanine aminotransferase<\/td>\n<\/tr>\n<tr>\n<td>AST<\/td>\n<td>aspartate aminotransferase<\/td>\n<\/tr>\n<tr>\n<td>BUN<\/td>\n<td>blood urea nitrogen<\/td>\n<\/tr>\n<tr>\n<td>CAT<\/td>\n<td>catalase<\/td>\n<\/tr>\n<tr>\n<td>CLP<\/td>\n<td>cecal ligation and puncture<\/td>\n<\/tr>\n<tr>\n<td>Cr<\/td>\n<td>creatinine<\/td>\n<\/tr>\n<tr>\n<td>GSH-Px<\/td>\n<td>glutathione peroxidase<\/td>\n<\/tr>\n<tr>\n<td>H<sub>2<\/sub><\/td>\n<td>hydrogen<\/td>\n<\/tr>\n<tr>\n<td>H<sub>2<\/sub>O<sub>2<\/sub><\/td>\n<td>hydrogen peroxide<\/td>\n<\/tr>\n<tr>\n<td>HMGB1<\/td>\n<td>high-mobility group box 1<\/td>\n<\/tr>\n<tr>\n<td>ICU<\/td>\n<td>intensive care units<\/td>\n<\/tr>\n<tr>\n<td>MODS<\/td>\n<td>multiple organ dysfunction syndrome<\/td>\n<\/tr>\n<tr>\n<td>NS<\/td>\n<td>normal saline<\/td>\n<\/tr>\n<tr>\n<td>\u2022OH<\/td>\n<td>hydroxyl radicals<\/td>\n<\/tr>\n<tr>\n<td>ROS<\/td>\n<td>reactive oxygen species<\/td>\n<\/tr>\n<tr>\n<td>SOD<\/td>\n<td>superoxide dismutase<\/td>\n<\/tr>\n<tr>\n<td>ZY<\/td>\n<td>zymosan<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/div>\n<\/div>\n<\/div>\n<div id=\"ref-list-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=\"ref-list-a.h.ctitle\">References<\/h2>\n<div class=\"ref-list-sec sec\" id=\"reference-list\">\n<div class=\"ref-cit-blk half_rhythm\" id=\"b1-etm-0-0-3231\">1. <span class=\"element-citation\">Martin GS, Mannino DM, Eaton S, Moss M. 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Severity of sepsis correlated with the elevation of serum high-mobility group box 1 in rats. <span><span class=\"ref-journal\">Chin Med J (Engl) <\/span>2009;<span class=\"ref-vol\">122<\/span>:449\u2013454.<\/span> [<a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/19302753\" ref=\"reftype=pubmed&amp;article-id=4887775&amp;issue-id=270295&amp;journal-id=1895&amp;FROM=Article%7CCitationRef&amp;TO=Entrez%7CPubMed%7CRecord\">PubMed<\/a>] <span class=\"nowrap\">[<a href=\"https:\/\/scholar.google.com\/scholar_lookup?journal=Chin+Med+J+(Engl)&amp;title=Severity+of+sepsis+correlated+with+the+elevation+of+serum+high-mobility+group+box+1+in+rats&amp;author=LC+Hou&amp;author=MZ+Qin&amp;author=LN+Zheng&amp;author=Y+Lu&amp;author=Q+Wang&amp;volume=122&amp;publication_year=2009&amp;pages=449-454&amp;pmid=19302753&amp;\" target=\"_blank\" rel=\"noopener noreferrer\" ref=\"reftype=other&amp;article-id=4887775&amp;issue-id=270295&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.f.b.utitle\">Abstract<\/a><\/li>\n<li><a href=\"#sec-a.g.atitle\">Introduction<\/a><\/li>\n<li><a href=\"#sec-a.g.btitle\">Materials and methods<\/a><\/li>\n<li><a href=\"#sec-a.g.ctitle\">Results<\/a><\/li>\n<li><a href=\"#sec-a.g.dtitle\">Discussion<\/a><\/li>\n<li><a href=\"#ack-a.h.atitle\">Acknowledgements<\/a><\/li>\n<li><a href=\"#glossary-a.h.btitle\">Glossary<\/a><\/li>\n<li><a href=\"#ref-list-a.h.ctitle\">References<\/a><\/li>\n<\/ul>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Combination therapy of molecular hydrogen and hyperoxia improves survival rate and organ damage in a zymosan-induced generalized inflammation model<\/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":[843],"body-organ":[1022],"applications":[679],"test_subjects":[1518],"report-topic":[1329],"class_list":["post-27309","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-hydrogen-health","disease-sepsis-2","body-organ-whole-body-2","applications-inhalation-2","test_subjects-mouse-2","report-topic-multiple-organ-dysfunction-syndrome-mods-2"],"acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v28.1 - 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