Research Of The Expression Of HMGB-1 MRNA And Protein In Hepatic Ischemic Reperfusion Injury And The Relationship With Apoptosis

Objective:To detect the apoptosis and the expression of HMGB-1 mRNA and HMGB-1 protein in hepatic tissue during the course of ischemic reperfusion, then discuss the response of hepatic tissue to ischemic reperfusion and the relationship between HMGB-1 protein and apoptosis. To supply the theory of effective pretreatment, apoptosis depression and lessening hepatic ischemic reperfusion injury.Methods:1,Sample collection: We Selected 36 patients with hepatocellular carcinoma who take liver resection operation at the First Affiliated Hospital of Guangxi Medical University from March 2007 to March 2009, all of them no line chemotherapy and embolization before operative, patients in both use of Pringle's France blocked portal.Hepatic tissue(3gram)was taken around incisal before the blockage of porta hepatis, the open of porta hepatis and before the closure of abdominal cavity respectively, to take the 3g hepatic tissue to bisection two portions, the one portion was fixed with 4% Polyoxymethylene, then were embedded in paraffin wax; the other portion was conserved in -70℃freezer, then were used to abstraction common RNA. 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 specimens we taken was consent by the patients and also reviewed and approved by Hospital Ethics Committee.2,The expression of HMGB-1 mRNA was determined by reverse transcription polymerase chain reaction(RT-PCR).3,The hepatocellular apoptosis of the hepatic tissue was determined by terminal-deoynucleotidle transferase mediated d-UTP nick end labeling method.4,The expression of HMGB-1 protein in hepatic tissue was observed by immunohistochemical method.Results:1,The expression of HMGB-1 mRNA in the pre-blockage of porta hepatis group was 0.5554±0.0843, the open of porta hepatis group was 1.0703±0.1931, the pre-closure of abdominal cavity group was 1.4028±0.1931. Differences among these three groups had distinguished statistic significance(p<0.05). Compared with the open of porta hepatis group, the expression of HMGB- 1mRNA declined markedly in the pre-blockage of porta hepatis group. Compared with the open of porta hepatis group, the expression of HMGB-1 mRNA increased markedly in the pre-closure of abdominal cavity group.2,Hepatocyte Apoptosis Index in the pre-blockage of porta hepatic group was 2.89±0.274%, the open of porta hepatis group was 2.26±0.337%,the pre-closure of abdominal cavity group was 1.51±0.524%. Differences among these three groups had distinguished statistic significance(p<0.05). Compared with the pre-blockage of porta hepatis group, Apoptosis Index declined markedly in the open of porta hepatis group. Compared with the open of porta hepatis group, Apoptosis Index declined markedly in the pre-closure of abdominal cavity group.3,The expression of immunohistochemical staining film scores in the pre-blockage of porta hepatic group was 2.97±0.576,the open of porta hepatis group was6.51±1.354,the pre-closure of abdominal cavity group was 10.42±0.985.Differences among these three groups had distinguished statistic significance(p<0.05). Differences among these three groups had distinguished statistic significance(p<0.05).Compared with the open of porta hepatis group, the expression of immunohistochemical staining film scores increased markedly in the pre-closure of abdominal cavity group. Compared with the pre-blockage of porta hepatis group, the expression of immunohistochemical staining film scores increased markedly in the open of porta hepatis group.4,①In the pre-blockage of porta hepatis group, the correlation coefficient between HMGB-1 mRNA expression in hepatic tissue and the expression of HMGB-1 protein in hepatic tissue wasγ=0.905, p<0.01, the HMGB-1 mRNA expression has a high positive correlation with the expression of HMGB-1 protein in hepatic tissue.②In the pre-blockage of porta hepatis group, the expression of HMGB-1 mRNA has a high negative correlation with hepatocyte apoptosis in hepatic tissue, the correlation coefficient wasγ= - 0.831, p<0.01.③In the open of porta hepatis group, the correlation coefficient between HMGB-1mRNA expression in hepatic tissue and the expression of HMGB-1 protein in hepatic tissue wasγ=0.927, p<0.01, the HMGB-1 mRNA expression has a high positive correlation with the expression of HMGB-1 protein in hepatic tissue.④In the open of porta hepatis group, the expression of HMGB-1 mRNA has a high negative correlation with hepatocyte apoptosis in hepatic tissue, the correlation coefficient wasγ=- 0.878, p<0.01.⑤In the pre-closure of abdominal cavity group, the correlation coefficient between HMGB-1 mRNA expression in hepatic tissue and the expression of HMGB-1 protein in hepatic tissue wasγ=0.940, p<0.01, the HMGB-1 mRNA expression has a high positive correlation with the expression of HMGB-1 protein in hepatic tissue.⑥In the pre-closure of abdominal cavity group, the expression of HMGB-1 mRNA has a high negative correlation with hepatocyte apoptosis in hepatic tissue, the correlation coefficient wasγ= -0.932, p<0.01.Conclusion:1,There is little expression of HMGB-1 mRNA and HMGB-1 protein 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 HMGB-1 and HIRI.2,The expression of HMGB-1 mRNA and HMGB-1 protein in the open of porta heptis group increase, the AI decreases, there is inverse correlation between HMGB-1 and the AI. It suggests HMGB-1 has an effect on suppressing apoptosis.