The Mechanism And Protection Of Myocardial And Mitochondrion Injury After Total Liver Ischemia-reperfusion

Background and Objective:Liver transplantation and most liver resection are clinically normal and the surgical techniques are quite mature.However,the blood flow in the inferior vena cava,portal vein and hepatic artery need to be blocked during the operation,thus the complexity of surgical procedures results in total hepatic ischemia-reperfusion injury,which not only affects the function of the liver itself,but also damage the distant organs-cardiovascular system.In the experiment,we established the model of total rat liver ischemia-reperfusion, Firstly,we dynamically detect the changes of H₂S,TNF-α,MDA,Angâ…¡,ALD, CK-MB,LDH,the activity of myocardial CSE and the content of ALD,NF-kB. We detect the expression of myocardial CaNAβmRNA and ALDH2 mRNA, observe the changes of the myocardial tissue CaN Aβgene expression using RT-PCR,observe the pathological damage of the myocardial with light microscope and observe the pattern and structure changes of the myocardial. Secondly,we design to use the NaHS,MP,spironolactone and FK506 to interfere in,observing the same index as above.Lastly,we determine the RCR,P/O, membrane potential,SDH,Na⁺/K⁺ ATPase,Ca²⁺-ATPase of myocardial mitochondrion,to research the mechanism of myocardial and mitochondrion injury after amputation trauma.Method:We have established the model of total rat liver ischemia-reperfusion by surgery.In the first part,88 Wistar rats were divided into the control group,20min ischemia,2h,4h,6h,12h,24h,48h and 72h after 20min ischemia,.40 min ischemia,6h and 72h after 40min ischemia,there are eight rats in each class.In the second part,128 Wistar rats were divided into 16 groups: the control group,6h after 20min ischemia,6h and 72h after 40min ischemia NaHS group:6h after ischemia +NaHS 28μmol/kg ip;MP group:6h after ischemia+MP 30mg/kg ip;Spironolactone group:6h after ischemia+ spironolactone 20mg/kg intragastric administration for six days;FK506 group:6h after ischemia +FK506 0.1mg/kg ip,the responding medicine blank group(no blood vessel blocked),MP and Spironolactone group for 6h repufusion after 40 min ischemia,and the MP and Spironolactone group for 72h repufusion after 40 min ischemia.In the last part,80 Wistar rats were divided into 10 groups as 6h after 20 min ischemia of the second part,observed the function of mitochondrion.Result:The results of the fist part were as follows:All variables were compared with the control group.The levels of TNF-α,MDA in plasma and the levels of MDA in myocardium were significantly increased after liver ischemia (P＜0.05),reached a peak 6h and 12h after reperfusion group,and then decreased slowly.The levels of H₂S in plasma increased significantly at 2h after ischemia and reached a peak 6h after reperfusion,then reached baseline 72h.The myocardial level of CSE was markedly decreased and reached the lowest in 12h group.The results of the second part were as follows:All variables were compared with the 6h after reperfusion group,the levels of MDA,TNF-αNF-kB and ICAM-1 in plasma and the myocardial in four group were significantly lower.The myocardial CSE activity in NaHS and MP group was significantly lower.The plasma H₂S were significantly higher in NaHS group.The results of the last part were as follows:compared to the control group,respiratory function of mitochondrion was broken-down and the levels of RCR,P/O,Na⁺/K⁺ ATP,SDH and membrane potential were significantly lower,Ca²⁺-ATP were significantly higher.Compared to the 6h after reperfusion group,respiratory function of mitochondrion was improved and the levels of RCR,P/O,Na⁺/K⁺ ATP, SDH and membrane potential were higher in NaHS,FK506,MP groups.The levels of Ca²⁺-ATP were decreased in the above groups.The levels of RCR,ATP and membrane potential in spironolactone group were higher,and the levels of Ca²⁺-ATP were decreased in spironolactone group.Conclution:Total liver ischemia and reperfusion can induce remote myocardial and mitochondrion injury.The mechanism may be involved in the activating of neutrophil and inflammatory cytokines,excess oxidative stress. Down-regulation of endogenous H₂S/CSE system and CaN signal transduction also participate in the myocardial and mitochondrion injury after Total liver ischemia and reperfusion.Exogenous H₂S,FK506 can protect myocardial injury after reperfusion trauma.