川芎嗪对血管紧张素Ⅱ诱导的大鼠肝星状细胞转化生长因子-β1/Smads通路的影响

Effects of Tetramethylpyrazine and angiotensin Ⅱ on TGF-β1/Smads signaling pathway in rat hepatic stellate cells

摘要:

目的  通过研究川芎嗪对体外培养的血管紧张素Ⅱ诱导的肝星状细胞（HSC）转化生长因子-β1（TGF-β1）/Smads通路的作用，探讨其治疗肝纤维化的机制。方法  对大鼠HSC使用血管紧张素Ⅱ（1×10-5 mol/L）及血管紧张素Ⅱ（1×10-5 mol/L）联合不同浓度的川芎嗪（0.04、0.20和1.00 mg/ml）干预48 h，活细胞计数法（CCK-8）检测各组细胞增殖水平的变化，实时荧光定量聚合酶链反应及Western blot检测各组细胞TGF-β1、Smad2、Smad4的mRNA及蛋白表达。结果  血管紧张素Ⅱ明显促进HSC的增殖，同时上调该细胞TGF-β1、Smad2、Smad4的mRNA及蛋白表达（P <0.01），而血管紧张素Ⅱ对HSC的诱导作用可被川芎嗪抑制（P <0.01），并呈现浓度依耐性。结论  川芎嗪抗肝纤维化的机制之一可能是通过调控肝星状细胞TGF-β1/ Smads通路实现的。

关键词:

肝星状细胞;转化生长因子-&beta;1/Smads;川芎嗪;血管紧张素Ⅱ

Abstract:

Objective To investigate the effects of Tetramethylpyrazine and angiotensin Ⅱ on transforming growth factor-β1 (TGF-β1) / Smads signaling pathway in rat hepatic stellate cells (HSCs). Methods HSCs were cultured, and then treated by angiotensin Ⅱ (1×10-5 mol/L) or angiotensin Ⅱ (1×10-5 mol/L) combined with different concentrations of Tetramethylpyrazine (0.04, 0.20 and 2.00 mg/ml). The treated cells and control groups were incubated for 48 hours. Cell proliferation was assayed by CCK-8. Relative mRNA expressions of TGF-β1, Smad2 and Smad4 were assayed by quantitative real-time polymerase chain reaction. Relative protein expressions of TGF-β1, Smad2 and Smad4 were investigated by Western blot. Results Angiotensin Ⅱ significantly promoted HSC proliferation, significantly up-regulated both the mRNA and protein expressions of TGF-β1, Smad2 and Smad4 in HSCs (P < 0.01). The effects of angiotensin Ⅱ in HSCs were significantly inhibited by Tetramethylpyrazine in a dose-dependent manner (P < 0.01). Conclusions This study suggested that the antiliver fibrosis effects of Tetramethylpyrazine maybe connect with modulation of TGF-β1/Smads signaling pathway.

Key words:hepatic stellate cell; TGF-β1/Smads; Tetramethylpyrazine; angiotensin Ⅱ