线粒体转录因子A对梗阻性黄疸大鼠肝功能的保护作用.docVIP

线粒体转录因子A对梗阻性黄疸大鼠肝功能的保护作用 林 恒，唐 春，吴 乔，冯春林，张玉君，别 平 （第三军医大学西南医院全军肝胆外科研究所，重庆 400038） 摘 要：目的 初步探讨线粒体转录因子A(mitochondrial transcription factor A，TFAM)在重组人肝再生增强因子(rhALR)对梗阻性黄疸大鼠肝功能保护过程中的作用。 方法 首先体外合成靶向TFAM的有效慢病毒载体，然后建立梗阻性黄疸大鼠动物模型，并通过门静脉将慢病毒注入大鼠肝组织进行靶向TFAM活体RNA干扰实验。然后通过RT-PCR和Western blot检测目的基因表达量的改变并观察大鼠肝脏病理改变、肝脏功能改变及rhALR保护作用的变化情况。结果 体外靶向TFAM慢病毒载体构建成功。通过门静脉注入后能特异、有效的感染肝组织细胞。RT－PCR和Western blot检测结果显示进行活体RNAi的大鼠肝组织中TFAM的表达明显减弱；且这一组大鼠肝脏病理改变最为明显，肝功能损害最严重，rhALR的肝功能保护作用也随着消失。结论 阻断目的基因TFAM的表达使得rhALR对梗阻性黄疸大鼠肝功能的保护作用消失；TFAM在rhALR保护梗阻性黄疸大鼠肝功能过程中具有重要作用。 关键词：线粒体转录因子A；体内，RNA干扰；重组人肝再生增强因子；梗阻性黄疸；慢病毒载体 中图法分类号： 文献标识码：A Protective effects of mitochondrial transcription factor A on hepatic function in rats with obstructive jaundice. Lin Heng, Tang Chun, Wu Qiao, Feng Chun-lin, Zhang Yu-jun, Bie Ping（Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing 400038, China） Abstract：Objective To explore the role of mitochondrial transcription factor A (TFAM) in protecting the hepatic function of rats with obstructive jaundice by recombinant human augmenter of liver regeneration (rhALR). Methods The effective lentivirus-siRNA vector system targeting TFAM was successfully constructed in vitro. Then we established obstructive jaundice rat model and the lentivirus-siRNA vector was infused into liver through portal vein for RNAi in vivo. RT-PCR and Western blot analysis were used to detect the change of TFAM expression, liver pathology, hepatic function and the effect of rhALR. Results Liver cells were specially and effectively transfected by lentivirus-siRNA vector. The results of RT-PCR and WB showed that TFAM expression was obviously suppressed. The knockdown of target gene led to obviously changed hepatic pathology andseverely damaged hepatic function. The protective effect of rhALR also disappeared. Conclusion The protective effect of rhALR on hepatic function disappeared after the suppression o