Bcl-2基因修饰的肝细胞移植对肝脏缺血再灌注损伤的影响

Attenuation of hepatic ischemia/reperfusion injury by transplantation of Bcl-2 gene modified hepatocytes

目的观察人Bcl-2(hBcl-2)基因修饰的肝细胞移植对肝脏缺血再灌注损伤的影响。方法将包含hBcl-2基因阅读框架(0·7kb)的核苷酸亚克隆至Psectag2A载体,脂质体转染balb/c小鼠胎肝细胞(BNL.CL2),经门静脉移植入balb/c小鼠(2×106细胞)。移植72h后左肝后叶常温下缺血45min后再灌注8h,比较测定凋亡细胞,血清ALT、AST、LDH,以及组织学的改变。结果转染hBcl-2基因的balb/c小鼠在肝脏缺血再灌注后ALT(P<0·05或0·01)、AST(P<0·05)、LDH(P<0·05)、凋亡细胞(P<0·05)均较对照组显著降低,组织学改变减轻。进行缺血再灌注的各组细胞损伤均以细胞凋亡为主(P<0·05或0·01)。结论hBcl-2基因修饰的肝细胞移植对肝脏缺血再灌注损伤具有保护作用,主要通过抑制缺血再灌注后的细胞凋亡。

Objective To investigate the effect of transplantation of Bcl-2 gene modified hepatocytes on hepatic ischemia/reperfusion injury. Methods After the balb/c mouse fetal hepatocytes (BNL.CL2) were transfected with Psectag2A encoding 0.7 kb human Bcl-2 gene open reading frame by Lipofectin, they were transplanted into the liver of balb/c mice by direct injection through the portal vein. The mice transplanted with Psectag2A BNL.CL2, BNL.CL2 or PBS were employed to serve as the controls. The serum levels of ALT, AST and LDH and apoptotic rate were determined by Annexin V and histological examination after lobus ischemia for 45 min followed by reperfusion for 8 h. Results The serum levels of ALT, AST and LDH and apoptotic rate were significantly lower and histological damage slighter in the mice receiving transplantation of human Bcl-2 gene modified BNL.CL2 than in the controls. Conclusions Transplantation of human Bcl-2 gene modified hepatocytes can ameliorate hepatic ischemia/reperfusion injury.