C/EBP-α 基因在大鼠肝星状细胞株 HSC-T6 自噬中的作用

Effect of C/EBP-α on autophagy in rat hepatic stellate cell line HSC-T6

摘要:

目的 探讨CCAT 增强子结合蛋白α（C/EBP-α）在肝星状细胞（HSCs）自噬过程中是否发挥作用。 方法 构建携带C/EBP-α 的慢病毒载体Lenti-C/EBP-α，感染大鼠肝星状细胞株HSC-T6，Western blot 检 测自噬相关蛋白 LC3（ 微管相关蛋白 1 轻链 3）表达；采用自噬激活剂 Rapamycin处理 HSC-T6，Western blot 检测C/EBP-α 和LC3 的表达；将HSC-T6 经Lenti-C/EBP-α 及自噬激活剂Rapamycin 共处理，Western blot 检测C/EBP-α 和LC3 的改变。结果 HSC-T6 感染Lenti-C/EBP-α 后，自噬活性降低，LC3 表达下降； Rapamycin 处理HSC-T6 后，自噬活性增高，C/EBP-α 表达下降，LC3 表达增多；HSC-T6 经Lenti-C/ EBP-α 及自噬激活剂Rapamycin 共处理，与单纯使用Rapamycin 比较，C/EBP-α 表达升高，LC3 激活减少， 抑制HSCs 激活。结论 C/EBP-α 可以抑制自噬激活，从而抑制HSCs 激活，为C/EBP-α 抗肝纤维化提出 了新思路。

Abstract:

Abstract: Objective To investigate the role of CCAT enhancer binding protein α (C/EBP-α) in autophagy of hepatic stellate cells (HSCs). Methods C/EBP-α Lentiviral vector was constructed. HSC-T6 was treated with C/ EBP-α Lentiviral vector, autophagy activator Rapamycin or C/EBP-α Lentiviral vector plus Rapamycin. Autophagyrelated protein LC3 and C/EBP-α were detected with Western blotting. Results Treatment of Lenti-C/EBP-α induced decreased expression of LC3 in HSC-T6 while Rapamycin upregulated levels of LC3 and downregulated expression of C/EBP-α.Co-incubation of Lenti-C/EBP-α and Rapamycin suggested that C/EBP-α expression was increased and LC3 expression was decreased when compared with Rapamycin only group. Conclusion C/EBP-α inhibits activation of HSC-T6 inhibition of autophagy, which may be a novel approach for anti-hepatic fibrosis.