南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (05): 561-.doi: 10.12122/j.issn.1673-4254.2019.05.10

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PPAR-γ调控的自噬通路在染料木黄酮抑制肝星状细胞激活过程中的作用

刘希鹏，张美芳，张海峰，赵安达，孙娟，唐雯   

  • 出版日期:2019-05-20 发布日期:2019-05-20

Role of PPAR-γ-regulated autophagy in genistein-induced inhibition of hepatic stellate cell activation

  • Online:2019-05-20 Published:2019-05-20

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摘要： 目的探讨染料木黄酮在体外抑制肝星状细胞（HSC）活化的效应以及PPAR-γ调控的自噬通路在其中发挥的作用。方法 将体外培养的HSC细胞分别给予不同浓度的染料木黄酮作用48 h后，用Western blot法测定HSC激活标志蛋白α-SMA、α1（I） collagen的表达以及自噬相关蛋白LC3Ⅱ\p62和PPAR-γ的表达；利用自噬抑制剂和PPAR-γ抑制剂验证自噬在染料木黄酮抑制 HSC激活过程中的作用以及PPAR-γ对自噬的调控作用。结果染料木黄酮作用于HSC细胞后，HSC活化标志蛋白α-SMA、α1 （I）collagen的水平明显降低（P<0.05），而自噬相关蛋白LC3Ⅱ的表达明显升高、泛素结合蛋白p62水平明显降低，同时PPAR-γ 表达明显增加（P<0.05）；与单纯染料木黄酮刺激组相比，用3-MA处理后HSC活化标志蛋白α-SMA、α1（I）collagen的表达明显 升高；并且加入PPAR-γ抑制剂后，自噬相关蛋白LC3Ⅱ的表达明显降低、泛素结合蛋白p62 水平明显增加（P<0.05）。结论 PPAR-γ调控的自噬通路在染料木黄酮抑制HSC细胞激活的过程中发挥着重要作用。

Abstract: Objective To investigate the inhibitory effect of genistein on activation of hepatic stellate cells (HSCs) in vitro and the role of the autophagy pathway regulated by PPAR-γ in mediating this effect. Methods Cultured HSC-T6 cells were exposed to different concentrations of genistein for 48 h, and HSC activation was verified by detecting the expressions of α-SMA and α1(I) collagen; autophagy activation in the cells was determined by detecting the expressions of LC3-II and p62 using Western blotting. The autophagy inhibitor 3-MA was used to confirm the role of autophagy in genistein- induced inhibition of HSC activation. A PPAR-γ inhibitor was used to explore the role of PPAR-γ in activating autophagy in the HSCs. Results Genistein at concentrations of 5 and 50 μmol/L significantly inhibited the expressions of α-SMA and α1(I) collagen (P<0.05), markedly upregulated the expressions of PPAR-γ and the autophagy-related protein LC3-II (P<0.05) and significantly down-regulated the expression of the ubiqutin-binding protein p62 (P<0.05) in HSC-T6 cells. The cells pretreated with 3-MA prior to genistein treatment showed significantly increased protein expressions of α-SMA and α1(I) collagen compared with the cells treated with genistein only (P<0.05). Treatment with the PPAR-γ inhibitor obviously lowered the expression of LC3-II and enhanced the expression p62 in genistein-treated HSC-T6 cells, suggesting the activation of the autophagy pathway. Conclusion PPAR-γ- regulated autophagy plays an important role in mediating genistein-induced inhibition of HSC activation in vitro.