Clinical Study Of The Expression Of HSP70 MRNA And Protein In Hepatic Ischemic Reperfusion Injury And The Relationship With Apoptosis

Objective:To detect the apoptosis and the expression of HSP70 mRNA and protein in hepatic tissue during the course of ischemic reperfusion,then discuss the response of hepatic tissue to ischemic reperfusion and the relationship between HSP70 and apoptosis.To supply the theory of effective pretreatment,apoptosis depression and lessening hepatic ischemic reperfusion injury.Methods:1,Sample collection:The blood sample from peripheral vein was obtained in pre-operation,the end of operative,the first day,the fourth day and the seventh day after operation.The blood sample was sent to centrifuge to obtain serum.Hepatic tissue(10 gram)was taken around incisal margin before the blockage of porta hepatis,the open of porta hepatis and before the closure of abdominal cavity respectively,then were embedded in paraffin wax.2,In situ hybridization was used to detect the expression of HSP70 mRNA. Immunohistochemical S-P method was used to detect the expression of HSP70. Terminal deoxynucleotidy transferase mediated dUTP nick end labeling was used to detect the apoptosis of hepatic tissue.Enzyme linked immunosorbent assay was used to detect the expression of HSP70 in serum.Automatic biochemistry analyzer was used to detect liver function.Result:1,The number of positive cell of HSP70 mRNA expression in the pre-blockage of porta hepatis group was 1.90±0.84,the open of porta hepatis group was 24.4±5.7,the pre-closure of abdominal cavity group was 35.7±5.7. Differences among these three groups were significant(P＜0.05).2,The number of positive cell of HSP70 expression in the pre-blockage of porta hepatis group was O,the open of porta hepatis group was 19.8±4.4,the pre-closure of abdominal cavity group was 29.9±6.2.Differences among these three groups were significant(P＜0.05).The difference of gray scale of HSP70 in the pre-blockage of porta hepatis group was 0.1609±0.1558,the open of porta hepatis group was 0.3409±0.3694,the pre-closure of abdominal cavity group was 0.5896±0.1802.Differences among these three groups were significant (P＜0.05).3,The AI(Apoptosis Index)of hepatic tissue in the pre-blockage of porta hepatis group was(11.4±2.4)‰,the open of porta hepatis group was(9.8±1.7)‰,the pre-closure of abdominal cavity group was(8.6±1.6)‰.The AI of hepatic tissue in the pre-blockage of porta hepatis group was more than the open of porta hepatis group(P＜0.05)and the pre-closure of abdominal cavity group(P＜0.05).The AI of hepatic tissue in the open of porta hepatis group was more than the pre-closure of abdominal cavity group(P＜0.05).4,The optical density of serum HSP70 in the pre-blockage of porta hepatis group was 1.61±0.24,the pre-closure of abdominal cavity group was 1.58±0.21,the group of the 1^stday after operation was 1.66±0.26,the group of the 4^thday after operation was 1.74±0.27,the group of the 7^thday after operation 1.68±0.24. Differences among these three groups were significant(P＜0.05).5,The differences of ALT and AST among these four groups were significant(P＜0.05).Conclusion:1.There is little expression of HSP70 mRNA and HSP70 in the normal hepatic tissue.There is high expression in hepatic ischemic reperfusion injury(HIRI).The tendency rises,it suggests there is some relationship between HSP70 and HIRI.2.The expression of HSP70 mRNA and HSP70 in the open of porta heptis group increase,the AI decreases,there is inverse correlation between HSP70 and the AI.It suggests HSP70 has an effect on suppressing apoptosis.3.HSP70 is a protective factor which has an effect on recovery of organism and liver function.