Parkin基因对大鼠肝脏缺血再灌注损伤细胞自噬的影响

Autophagy Effect of Parkin Gene in Hepatic Ischemia Reperfusion Injury in Rats

目的探讨帕金森相关蛋白Parkin基因在肝脏缺血再灌注损伤中对细胞自噬的作用。方法采用肝门阻断法建立大鼠肝脏缺血再灌注模型,实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)和Western blot法分别检测Parkin基因及蛋白表达水平在缺血再灌注前后的变化,同时检测缺血再灌注前后细胞自噬关键蛋白LC3的表达变化;构建靶向Parkin基因的RNA干扰质粒,检测下调Parkin基因表达后对缺血再灌注损伤细胞自噬的影响。结果肝脏缺血再灌注损伤后,Parkin蛋白的表达水平显著增加,同时肝细胞自噬发生显著增加。下调Parkin基因表达后,肝细胞自噬显著降低(t=11.94,P=0.003)。结论 Parkin基因能够有效诱导肝脏缺血再灌注损伤中的细胞自噬的发生,是肝脏缺血再灌注损伤的保护性基因。

Objective To investigate the autophagy effects of Parkinson related protein Parkin gene in hepatic is- chemia reperfusion injury. Methods The rat model with hepatic ischemia reperfusion was established by hepatic portal occlusion. The changes of Parkin gene and protein expressions were detected by quantitative real-time polymerase chain reaction （qRT-PCR） and Western blot before and after ischemia reperfusion. Meanwhile, the expressions of cell autoph- agy key protein LC3 were detected before and after ischemia reperfusion. RNA interference plasmids of targeting Parkin gene were built, and autophagy effects of hepatic ischemia-reperfusion injury after down-regulated Parkin gene expres- s{ons were investigated. Results The expression of Parkin protein and hepatocytes autophagy significantly increased after hepatic ischemia reperfusion injury. After Parkin gene was down-regulated, the occurs of autophagy in liver cells reduced （t = 11.94, P = 0. 003）. Canclusion Parkin gene can effectively induce cell autophagy of hepatic isehemia-reperfusion in- jury, and it is a protective gene for liver ischemia reperfusion injury.