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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。
Objective To investigate the role of mitochondrial transcription factor A (TFAM) in the protection of liver function in rats with obstructive jaundice by recombinant human augmenter of liver regeneration (rhALR). Methods The effective lentiviral vector targeting TFAM was synthesized in vitro. Then an animal model of obstructive jaundice was established. The lentivirus was injected into rat liver tissue through portal vein to perform RNA interference targeting TFAM. The expression of target gene was detected by RT-PCR and Western blot. The changes of hepatic pathological changes, liver function and rhALR protective effect were observed. Results The in vitro targeting TFAM lentiviral vector was successfully constructed. Through the portal vein after injection can be specific and effective infection of liver tissue cells. The results of RT-PCR and Western blot showed that the expression of TFAM in liver tissue of rats treated with live RNAi was significantly weakened. The pathological changes of liver in this group were the most obvious, and the liver damage was most serious. The hepatic function of rhALR was also protected Disappear Conclusion Blocking the expression of target gene TFAM results in the loss of protective effect of rhALR on liver function in rats with obstructive jaundice. TFAM plays an important role in protecting hepatic function of obstructive jaundice rats by rhALR.