肝缺血再灌注后HSP70 mRNA的表达及其与凋亡关系的临床研究

Clinical study on expression of HSP70 mRNA in hepatic ischemic reperfusion injury and its relationship with apoptosis

目的通过检测肝组织在缺血再灌注后HSP70 mRNA的表达及细胞凋亡情况,探讨肝组织对缺血再灌注的反应及HSP70mRNA与肝细胞凋亡之间的关系。为选用有效的预处理方法、抑制细胞凋亡、减轻肝脏缺血再灌注损伤提供理论基础。方法术中在肝门阻断前、肝门阻断后开放时和手术结束关腹前各取切缘旁肝组织10g。原位杂交法检测肝组织中HSP70 mRNA的表达;脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测肝细胞的凋亡情况。结果①肝门阻断前肝组织HSP70 mRNA在每高倍视野下阻性细胞数为1.90±0.84;肝门开放时为24.4±5.7;关腹前为35.7±5.7,3组阳性细胞数比较差异有统计学意义(P<0.05);两两比较差异有统计学意义(P<0.05)。②肝门阻断前肝细胞凋亡指数为(11.4±2.4)‰;肝门开放时为(9.8±1.7)‰;关腹前为(8.6±1.6)‰。肝门阻断前肝细胞的凋亡指数高于肝门开放时肝细胞的凋亡指数(P<0.05)、高于关腹前肝细胞的凋亡指数(P<0.05);肝门开放时肝细胞的凋亡指数高于关腹前肝细胞的凋亡指数(P<0.05)。③肝组织中HSP70 mRNA的表达与肝细胞凋亡指数呈负相关。结论①在正常肝组织中几乎无HSP70 mRNA的表达,术中肝缺血再灌注后的肝组织中HSP70 mRNA有较高的表达,且呈上升趋势,提示HSP70与HIRI的发生和发展有一定关系。②肝门阻断开放后肝组织HSP70 mRNA的表达增加,肝细胞凋亡指数减少,两者呈负相关。提示HSP70具有一定的抑制细胞凋亡作用,HSP70作为一种保护性因素对机体及肝功能的恢复起到一定作用。

Objective To detect the response of hepatic tissue to ischemic reperfusion and the relationship between the expression of HSP70 mRNA and apoptosis. And then to provide a theoretical basis for selecting effective pretreatment method, inhibiting apoptosis and reducing the hepatic injury by ischemic reperfusion. Methods liver tissue （ 10g for each） was taken around incisal margin before the blockage of porta hepatis, the open of porta hepatis and before the closure of abdominal cavity, respectively. In situ hybridization was used to detect the expression of HSP70 mRNA. Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling was used to detect the apoptosis in hepatic tissue. Results （1）The numbers of positive cell of HSP70 mRNA expression in the pre-hlockage of porta hepatis group, the open of porta hepatis group and the pre-closure of abdominal cavity group was 1.90 ± 0. 84, 24.4 ± 5.7 and 35.7 ± 5.7 respectively. Differences among these three groups were significant （P 〈0.05）. （2）The AI （apoptosis index） of hepatic tissue in the pre-blockage of porta hepatis group, the open of porta hepatis group and the pre-closure of abdominal cavity group was （ 11.4 ± 2.4 ） ‰, （ 9. 8 ± 1.7 ） ‰ and （ 8. 6 ± 1.6 ） ‰ respectively. AI in the pre-blockage of porta hepatis group was more than that in the open of porta hepatis group and the preclosure of abdominal cavity group （ P 〈 0.05 ）. AI in the open of porta hepatis group was more than that in the pre-closure of abdominal cavity group （ P 〈 0. 05 ）. Conclusions （1）There is little expression of HSP70 mRNA in the normal hepatic tissue, while there is high expression in hepatic ischemic reperfusion injury （HIRI）. It suggests that there is some relationship between HSP70 and HIRI. （2）In the open of porta heptis group, there is a negative correlation between the increased expression of HSP70 mRNA and the decreased of AI, which suggests HSP70 has an effect on suppressing apoptosis.