The Study Of Hydrogen-rich Water On Myocardial Ischemia Reperfusion Injury In Rats

Objective:To observe the effects of hydrogen-rich water on myocardial cell apoptosis and inflammatory factors in isolated rat.To probe into the protective effect of hydrogen rich water on myocardial ischemia reperfusion injury in isolated rat and its mechanism.Methods:Wistar 48 rats were randomly divided into the control group?24?and the hydrogen-rich water group?24?.The two groups were randomly divided into pre ischemic period,ischemic period and ischemia-reperfusion period.The heart was quickly removed from the rat after the rats were anesthetized,and the Langendorff model was set up.The control group was perfused with K-R solution?95%O₂+5%CO₂,7.85 kPa?,and the hydrogen-rich water group was perfused with K-R and hydrogen rich water?0.6 mmol/L,pH 7.3,95%O₂+5%CO₂,7.85kPa?.The isolated heart was treated by retrograde perfusion of 10 min,20 min at normal temperature and 20 min for reperfusion.Biochemical method was used to detect the activity of lactate dehydrogenase?LDH?and creatine kinase?CK?in myocardium.The level of tumor necrosis factor-??TNF-??and interleukin-1??IL-1??in myocardial tissue was detected by double anti sandwich ELISA method,Western Blot was used to detect the expression of high mobility group protein B1?HMGB1?and transcription factor NF-?B protein,Hematoxylin eosin?HE?staining and observation of pathological changes of myocardial tissue under microscope.The expression levels of cysteine protease3?Caspase3?,B lymphocyte-2?Bcl-2?and B related-2 X protein?Bax protein?were detected by immunohistochemistry.Results:1.Control group during ischemia and ischemia reperfusion myocardial LDH?CK activity were higher than pre ischemia?P<0.05?have statistical sense;The LDH?CK activity of myocardium was no significant difference and was not significantin in each phase of hydrogen-rich water group;Compared with the ischemia reperfusion period,the LDH?CK activity of the hydrogen-rich water group was significantly lower than that of the control group?P<0.05?have statistical sense.2.In the control group,comparison of ischemia,reperfusion and pre ischemiaTNF-??IL-1?were significantly increased?P<0.05?was statistically significant,comparison of ischemic reperfusion period and the ischemic period TNF-??IL-1?were significantly increased?P<0.05?was statistically significant;In the hydrogen-rich water group,comparison of ischemia,reperfusion and pre ischemiaTNF-??IL-1?were significantly increased?P<0.05?was statistically significant,comparison of ischemic reperfusion period and the ischemic period TNF-??IL-1?were significantly increased?P<0.05?was statistically significant;In the hydrogen-rich water group and the control group during the ischemia reperfusion period TNF-??IL-1?were significantly decreased?P<0.05?was statistically significant.3.In the control group,the ischemia reperfusion compared with the pre ischemic and ischemic,the content of HMGB1 protein and NF-?B protein increased?P<0.05?have statistical sense;In the hydrogen-rich water group,the ischemia reperfusion compared with the pre ischemic and ischemic,the content of HMGB1 protein and NF-?B protein increased?P<0.05?have statistical sense;The content of HMGB1 protein and NF-?B protein in the ischemic reperfusion period of the hydrogen-rich water group was significantly lower than that of the control group?P<0.05?have statistical sense.4.In the control group,the ischemic reperfusion was compared with the ischemic period,the expression level of Bcl-2 protein decreased?P<0.05?,and the expression level of Bax and Caspase3 protein increased?P<0.05?have statistical sense;There was no significant difference between the hydrogen-rich water group;The hydrogen-rich water group was compared with the control group,in the period of ischemia reperfusion,the expression of Bcl-2 protein was increased?P<0.05?,and the expression level of Bax and Caspase3 protein decreased?P<0.05?,have statistical sense.Conclusion:1.Hydrogen-rich water has an obvious protective effect on myocardial ischemia reperfusion injury in isolated rat heart.The expression of TNF-?and IL-1?was reduced and the expression of HMGB1 protein and NF-?B protein was downregulated,which are related to inhibition of inflammation.2.Hydrogen-rich water has an obvious protective effect on myocardial ischemia reperfusion injury in isolated rat heart.Cut down the expression of Bax and Caspase3 protein,up-regulation the expression of Bcl-2 protein,it is related to the inhibition of cardiac myocyte apoptosis.