Effect Of Hydrogen-rich Water On MAPK-ERK1/2 And JAK-STAT Signaling Pathways After Isolated Myocardial Ischemia Reperfusion Injury In Rats

Objective:To investigate the regulatory effects of hydrogen-rich water on MAPK-ERK1/2 and JAK-STAT signaling pathways in isolated myocardial ischemia-reperfusion injury in rats.Method:Rats were randomly divided into control group and hydrogen-rich water group.The two groups were randomly divided into pre-ischemic,ischemic and ischemia-reperfusion periods.Langendroff perfusion device was used.The control group was perfused with K-R solution.The hydrogen-rich water group was perfused with K-R solution and hydrogen-rich water.After the experiment,samples were taken and the apoptotic rate of myocardial cells in each group was detected by TUNEL method.Western Blot was used to detect the expression levels of Bcl-2,Bax,caspase3,ERK-ERK1/2 and p-ERK1/2.Immunohistochemical method was used to detect the expression of p-ERK1/2 protein in myocardial cells of each group.Protein chip technology was used to screen and analyze the active protein factors in myocardium of hydrogen-rich water group and control group after ischemia-reperfusion.KEGG pathway with difference was obtained by using the enrichment principle of Gene Ontology(GO).Western Blot and immunohistochemistry were used to detect the expression of JAK2,STAT1,STAT3,P-STAT1,P-JAK2 and P-STAT3 in myocardium after ischemia-reperfusion.Result:1.There was no apoptosis in the control group and the hydrogen-rich water group in the pre-ischemic period,and the apoptosis was obvious in the ischemic period.In the control group,myocardial cell apoptosis increased significantly during ischemia-reperfusion period compared with ischemia-reperfusion period(P < 0.01).Compared with the control group during ischemia-reperfusion period,myocardial cell apoptosis decreased significantly in the hydrogen-rich water group(P < 0.01).2.Bcl-2,Bax and caspase-3 protein expression levels were higher in the same group during ischemia-reperfusion period and ischemia-reperfusion period than in the pre-ischemia period(P < 0.05).The expression of Bcl-2 protein in hydrogen-rich water group was higher than that in ischemia-reperfusion period(P < 0.05).The expression of Bax in control group was higher than that in ischemia-reperfusion period(P < 0.05).Compared with the control group during ischemia-reperfusion period,the expression level of Bcl-2 protein increased(P < 0.05)and the expression level of Bax and caspase-3 protein decreased(P < 0.05)in the hydrogen-enriched water group during ischemia-reperfusion period.3.The expression of p-ERK1/2 / ERK1/2 increased during ischemia-reperfusion and ischemia compared with pre-ischemia in the same group(P < 0.05).The expression of p-ERK1/2 and p-ERK1/2 / ERK1/2 in hydrogen-rich water group was higher than that in ischemia-reperfusion period(P < 0.05).Compared with the control group,the expression of p-ERK1/2 and p-ERK1/2 / ERK1/2 increased in the hydrogen-rich water group during the ischemia-reperfusion period(P < 0.05).4.The results of protein chip screening showed that there were differences in 25 protein expressions between hydrogen-rich water group and control group after ischemia-reperfusion(P < 0.05).There were differences in 5 signal pathways,such as JAK-STAT signaling pathway,Cytokine-cytokine receptor interaction(P < 0.01).5.Western blot results showed that P-JAK2/JAK2,P-STAT3/STAT3 increased and P-STAT1/STAT1 decreased in the hydrogen-rich water group compared with the control group after ischemia-reperfusion(P < 0.01).6.Immunohistochemical results showed that the positive rate of P-JAK2 and P-STAT3 protein increased and the positive rate of P-STAT1 protein decreased in the hydrogen-rich water group after ischemia-reperfusion compared with the control group(P < 0.01).Conclusion:1.Hydrogen-rich water can inhibit apoptosis induced by ischemia-reperfusion injury in isolated heart.2.Hydrogen-rich water can alleviate myocardial ischemia-reperfusion injury and inhibit cardiomyocyte apoptosis by regulating MAPK-ERK1/2 signaling pathway.3.Hydrogen-rich water may alleviate ischemia-reperfusion injury by up-regulating the expression of JAK2-STAT3 signaling pathway.