To Study The Effect And Mechanism Of Hydrogen Gas On Oxidative Stress Induced By Myocardial Ischemical/Reperfusion Injury In Diabetes Rats

Objective: To study the effect and mechanism of hydrogen gas onoxidative stress which is induced by myocardial ischemical/reperfusion injury indiabetes rats. Methods:60male Sprague-Dawley （SD） rats (300-350g) wererandomly divided into six groups: non-diabetes and sham-operated(NS), non-diabetes and ischemical/reperfusion(NI/R), non-diabetes and hydrogen gasadministration (NH), diabetes and sham-operated(DS), diabetes andischemical/reperfusion(DI/R), diabetes and ischemical/reperfusion and hydrogengas administration (DI/RH),10rats one group. After intraperitoneal injection ofstreptozotocin (STZ), rats were STZ-treated rats with high glucose food for fourweeks, and then induced diabetes rats. Regional myocardial ischemia wasinduced by transient occlusion of the left anterior descending coronary artery.After30min of ischemia, we removed the tube followed by reperfusion for120min.5ml/kg H2(99.9%) gas was administered by intraperitoneal injectionduring reperfusion in DI/RH group. After120min of reperfusion, the heart wasremoved for histopathology, SOD、 MDA were detected in cardiac tissue, theexpression of8-OH-dG was detected by immunohistochemical procedures, Nrf2mRNA was detected using reverse transcription PCR.Results:1.SODactivity: Activity in the NI/R group were significantly declined compared to theNS group(P <0.01); however, Activity in the NH group were significantlyelevated compared to the NI/R group(P <0.01);Compared with the DS group,activity decreased obviously in the DI/R group(P <0.01); however,activity inthe DH group were significantly elevated compared to the DI/R group(P <0.01).The DS group compared with the NS group, the DI/R group comparedwith the NI/R group, the DH group compared with the NH group, their activityall have different levels of reduced (P <0.01).2. Determination of MDA content:MDA content in the NI/R group were significantly elevated compared to the NSgroup(P <0.01); however,content in the NH group were significantly declinedcompared to the NI/R group(P <0.01);Compared with the DS group, contentelevated obviously in the DI/R group(P <0.01); however,content in the DHgroup were significantly decreased compared to the DI/R group(P <0.01).TheDS group compared with the NS group, the DI/R group compared with the NI/Rgroup, the DH group compared with the NH group, their content all havedifferent levels of increase (P <0.01或P＜0.05).3.The percentage of positivecells： The percentage of positive cells in the NI/R group were significantlyelevated compared to the NS group(P <0.01); however,the percentage ofpositive cells in the NH group were significantly declined compared to the NI/Rgroup(P <0.01);Compared with the DS group, the percentage of positive cellselevated obviously in the DI/R group(P <0.01); however,the percentage ofpositive cells in the DH group were significantly decreased compared to theDI/R group(P <0.01).The DS group compared with the NS group, the DI/Rgroup compared with the NI/R group, the DH group compared with the NHgroup, the percentage of positive cells all have different levels of increase (P <0.01).4.Expression level of Nrf2mRNA: Expression level in the NI/R group were significantly elevated compared to the NS group(P <0.01);however,expression level in the NH group were significantly increasedcompared to the NI/R group(P <0.01);Compared with the DS group, expressionlevel increased obviously in the DI/R group(P <0.01); however,expressionlevel in the DH group were significantly elevated compared to the DI/R group(P<0.01). The DS group compared with the NS group, the DI/R group comparedwith the NI/R group, the DH group compared with the NH group, the percentageof positive cells all have different levels of increase (P <0.01).Conclusion：1.Diabetes can aggravated damage myocardial ischemia-reperfusion in rats, butintraperitoneal injection of hydrogen can effectively improve the diabeticmyocardial ischemia-reperfusion in rats.2.Intraperitoneal injection of hydrogencan effectively improve the diabetic myocardial ischemia-reperfusion in rats byimproving the activity of SOD and the level of resistance to oxidativestress.3.The mechanism of Hydrogen resistance to oxidative may be related tothe activation of endogenous Nrf2/ARE signaling pathway and regulation ofthe downstream related expression of antioxidant enzymes.