刘海防, 谢青. 内质网应激在肝细胞凋亡中的意义[J]. 蚌埠医学院学报, 2010, 35(8): 768-770.
    引用本文: 刘海防, 谢青. 内质网应激在肝细胞凋亡中的意义[J]. 蚌埠医学院学报, 2010, 35(8): 768-770.
    LIU Hai-fang, XIE Qing. Significance of endoplasmic reticulum stress in hepatocytes apoptosis[J]. Journal of Bengbu Medical College, 2010, 35(8): 768-770.
    Citation: LIU Hai-fang, XIE Qing. Significance of endoplasmic reticulum stress in hepatocytes apoptosis[J]. Journal of Bengbu Medical College, 2010, 35(8): 768-770.

    内质网应激在肝细胞凋亡中的意义

    Significance of endoplasmic reticulum stress in hepatocytes apoptosis

    • 摘要: 目的:探讨内质网应激在肝细胞凋亡中的意义。方法:原位二步灌流法分离并培养BALBC小鼠原代肝细胞,化学合成法制备抗半胱氨酸天冬氨酸蛋白酶-12小分子干扰RNA(caspase-12 siRNA),脂质体包裹转染小鼠原代肝细胞。毒胡萝卜内酯4μmol/L诱导肝细胞凋亡。RT-PCR、Western blot检测抑制效果;MTT比色试验检测细胞活力,反映细胞凋亡。结果:siRNA对小鼠原代肝细胞caspase-12 mRNA在浓度200nmol/L时抑制率为86.22%;对凋亡细胞procaspase-12抑制率为50.04%,与单纯诱导凋亡细胞差异有统计学意义(P<0.01);MTT比色试验检测发现,与单纯诱导凋亡细胞相比,转染siRNA后细胞活力增高51.65%,两者差异有统计学意义(P<0.05)。结论:抑制内质网应激介导细胞凋亡的特异酶caspase-12,能在一定程度上阻抑小鼠原代肝细胞凋亡的发生,内质网应激在肝细胞凋亡中具有重要意义。

       

      Abstract: Objective: To study the significance of endoplasmic reticulum stress in hepatocytes apoptosis.Methods: The BALB/c mouse primary hepatocytes were isolated by two-step in situ liver perfusion,and then induced apoptosis with 4 μmol/L thapsigargin.siRNA against caspase-12 were synthesized chemically,then transfected into mouse primary hepatocytes by liposomes.Efficacy of caspase-12-siRNA inhibition was analyzed with RT-PCR and Western blot;viability of hepatocyte following apoptosis was evaluated with MTT.Results: The inhibition of caspase-12 mRNA and procaspase-12 expression were 86.22% and 50.04% respectively with 200 nmol/L siRNA (P<0.01).Contrasted with apoptotic hepatocytes,the cell activity analyzed with MTT increased by 51.65% (P<0.05).Conclusions: The mouse primary hepatocyte apoptosis can be reduced when endoplasmic reticulum stress-induced caspase-12 is inhibited.Endoplasmic reticulum stress plays an important role in hepatocyte apoptosis.

       

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