过氧化物酶体增殖物PGC-1α对肝缺血再灌注损伤的保护作用研究

Protective effect of peroxisome proliferator receptor γ coactivator-1α on hepatic ischemia-reperfusion injury

目的探讨过氧化物酶体增殖物受体γ共激活因子-1α(PGC-1α)对肝缺血再灌注损伤的保护作用。方法构建大鼠肝缺血再灌注损伤模型,恢复灌注后12h,以蛋白免疫印迹(Western blot)检测PGC-1α表达情况,并检测肝脏活性氧、三磷酸腺苷(ATP)水平及血丙氨酸氨基转移酶(ALT)活性变化,评估肝功能情况。另一方面,构建PGC-1α慢病毒过表达载体,并在大鼠缺血再灌注前转染,于缺血再灌注后,再以Western blot检测PGC-1α表达,肝脏活性氧、ATP水平和血肝酶活性变化,评估PGC-1α表达对肝缺血再灌注损伤的保护作用。结果缺血再灌注肝脏PGC-1α表达较假手术组及对照组(未手术)明显降低(均P<0.05),同时肝脏活性氧族水平及ALT活性显著升高,而肝ATP产生减少(均P<0.05)。PGC-1α慢病毒过表达载体转染显著上调缺血再灌注肝脏PGC-1α表达,同时肝脏活性氧族水平及ALT活性较未转染组显著降低,而肝ATP产生增加(均P<0.05)。结论 PGC-1α表达有利于保护肝缺血再灌注损伤。

Objective To investigate the protective effect of peroxisome proliferator receptorγcoactivator（PGC）-1αon hepatic ischemia-reperfusion injury.Methods The rat model of hepatic ischemia-reperfusion injury was established.The expression of PGC-1αwas detected by Western blot after 12 hours of reperfusion.The changes of reactive oxygen species（ROS）,ATP level and serum liver enzyme activity were measured,and the liver function was evaluated.On the other hand,PGC-1α lentiviral overexpression vector was constructed and transfected in rat before ischemia-reperfusion.After ischemia-reperfusion,the expression of PGC-1α,liver ROS,ATP level were measured by Western blot to explore the protective role of PGC-1αin liver ischemia reperfusion injury.Results The expression of PGC-1α in ischemia-reperfusion liver was significantly lower than that in the control group（P〈0.05）.The level of ROS[（325.4±70.9）RLUvs.（108.5±25.2）RLU,P〈0.05],the ALT activity in serum[（367.8±82.7）U/L vs.（98.7±16.8）U/L,P〈0.05]in ischemia-reperfusion liver were increased than that in the control group,whereas liver ATP production was reduced[（6.1±3.7）pmol vs.（19.8±3.1）pmol,P〈0.05）].The expression of PGC-1α in the liver was significantly up-regulated by PGC-1α lentiviral overexpression vector（57.3 ± 21.3）U/L vs.（311.2±25.8）U/L,P〈0.05）,down-regulated ROS[（98.7±18.9）RLUvs.（300.2±45.6）RLU,P〈0.05]and serum glutamic-pyruvic transaminase[（105.3±21.3）U/L vs.（311.2±25.8）U/L,P〈0.05）],and increased liver ATP production[（17.3±3.1）pmol vs.（5.8±2.0）pmol,P〈0.05]in contrast to non-transfected rats.Conclusion PGC-1α contributes to protect liver ischemia reperfusion injury.