Song Ya-fang, Lü Zhi-gang, Xu Lie-ming
Department of Liver Cirrhosis, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
Zhong Xi Yi Jie He Xue Bao. 2012 Apr;10(4):454-61. doi: 10.3736/jcim20120415.
To investigate the effects of salvianolic acid B (SA-B) on extracellular signal-regulated kinase (ERK) signal transduction pathway activated by transforming growth factor-β1 (TGF-β1) in rat hepatic stellate cells (HSCs).
HSCs were isolated from male Sprague-Dawley rats by in situ perfusion and Nycodenz density-gradient centrifugation method. TGF-β1 and SA-B were directly added to the serum-free medium of HSCs. Total and phosphorylated ERK, MEK, Raf and α-smooth muscle actin (α-SMA) and type I collagen were assayed by Western blotting.
Phosphorylation of MEK in HSCs with or without TGF-β1 was inhibited by SA-B; however, phosphorylation of Raf in HSCs with or without TGF-β1 was not inhibited by SA-B. Expression of α-SMA in HSCs with TGF-β1 was inhibited by SA-B. Combination of SA-B and the inhibitors of ERK (PD98059) can effectively inhibit the expression of α-SMA. SA-B also inhibited synthesization of type I collagen in HSCs with or without TGF-β1.
The action point of SA-B inhibiting ERK signaling induced by TGF-β1 in HSCs is the inhibition of the phosphorylation of MEK. SA-B reduces the increase of expression of α-SMA and protein synthesization of type I collagen induced by TGF-β1 by means of inhibiting ERK signaling in activated HSCs of rats.
研究丹酚酸B(SA-B)对转化生长因子-β1(TGF-β1)激活的大鼠肝星状细胞(HSCs)细胞外信号调节激酶(ERK)信号转导通路的影响。
采用原位灌注和Nycodenz密度梯度离心法从雄性Sprague-Dawley大鼠分离HSCs。将TGF-β1和SA-B直接加入HSCs的无血清培养基中。通过蛋白质印迹法检测总ERK、磷酸化ERK、MEK、Raf以及α-平滑肌肌动蛋白(α-SMA)和I型胶原。
SA-B抑制有无TGF-β1刺激的HSCs中MEK的磷酸化;然而,SA-B不抑制有无TGF-β1刺激的HSCs中Raf的磷酸化。SA-B抑制有TGF-β1刺激的HSCs中α-SMA的表达。SA-B与ERK抑制剂(PD98059)联合可有效抑制α-SMA的表达。SA-B还抑制有无TGF-β1刺激的HSCs中I型胶原的合成。
SA-B抑制TGF-β1诱导的HSCs中ERK信号的作用点是抑制MEK的磷酸化。SA-B通过抑制大鼠活化HSCs中的ERK信号,减少TGF-β1诱导的α-SMA表达增加和I型胶原的蛋白质合成。