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

PAGE PAGE 6 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 The effect of RNA interference in vivo targetingProtective effects of mitochondrial transcription factor A on the beneficial effect of recombinant human augmenter of liver regeneration on hepatic function in rats with obstructive jaundice. Lin Heng, Tang Chun, Wu Qiao, Feng Chun-lin, Zhang Yu-jun, Bie Ping.（Institute of hHepatobiliary Surgery, Southwest hHospital, 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 Tthe change of TFAM expression, liver pathology, hepatic function and the effect of rhALR.were determined by RT-PCR and Western blot analysis.Liver pathology,hepatic function and the effect of rhALR were also analyzed. Results Liver cells were specially and effectively transfected by lentivirus-siRNA vector. The results of RT-PCR and WB showed that TFAM expression was obviou