Xue Dong-ying, Hong Jia-he, Xu Lie-ming
Institute of Liver Diseases, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.
Zhonghua Gan Zang Bing Za Zhi. 2004 Aug;12(8):471-4.
To investigate the inhibiting effect of salianic-acid B (SA-B) on mitogen-activated protein kinase (MAPK) Signaling in activated rat hepatic stellate cells (HSCs).
HSCs were isolated from normal rat by in situ perfusion and Nycodenz density-gradient centrifugation method. HSCs were primarily cultured on uncoated plastic for 7 days. Then cells were stimulated with 10ng/ml transforming growth factor-beta1 (TGF-beta1) after incubated with 10-6 M/L SA-B. The effects of SA-B on Extracellular-regulated kinase (ERK) expression and its phosphorylation. Transforming growth factor beta1 receptor I (TbetaR I) and transforming growth factor beta1 receptor II (TbetaR II) on HSCs, type I collagen expression in HSC Induced by TGF-beta1 were detected with western blot assay. Quantity of Type I collagen in the medium of HSCs was detected by ELISA. Matrix metalloproteinase 2, 9, 13 (MMP-2, MMP-9 and MMP-13) in the medium of HSCs was tested by Zymography.
The phosphorylation of ERK1/2 in HSCs with or without TGF-beta1 was inhibited by SA-B. The expression of TbetaR I and TbetaR II on HSCs can not be affected by SA-B. The synthesization of Type I collagen in HSCs was decreased by SA-B; The synthesization and secretion of type I collagen in HSCs with TGF-beta1 were reduced by SA-B too. SA-B had no effect on the activity of MMP-2 and MMP-13, but induced the activity of MMP-13.
SA-B inhibits ERK signaling induced by TGF-b1 in HSC. This inhibition has no association with the expression of TbetaR I and TbetaR II on HSCs. SA-B reduces the synthesization and secretion of Type I collagen in HSC by means of inhibiting TGF-beta1 signaling, which might be not related to the degrading activities of MMPs.
研究丹酚酸B（SA-B）对活化的大鼠肝星状细胞（HSCs）中丝裂原活化蛋白激酶（MAPK）信号通路的抑制作用。
采用原位灌注和Nycodenz密度梯度离心法从正常大鼠中分离HSCs。将HSCs在未包被的塑料培养皿上原代培养7天。然后在与10-6 M/L SA-B孵育后，用10ng/ml转化生长因子-β1（TGF-β1）刺激细胞。用蛋白质免疫印迹法检测SA-B对细胞外调节激酶（ERK）表达及其磷酸化、HSCs上转化生长因子β1受体I（TβR I）和转化生长因子β1受体II（TβR II）、TGF-β1诱导的HSCs中I型胶原表达的影响。用酶联免疫吸附测定法检测HSCs培养基中I型胶原的含量。用明胶酶谱法检测HSCs培养基中基质金属蛋白酶2、9、13（MMP-2、MMP-9和MMP-13）。
SA-B抑制了有无TGF-β1情况下HSCs中ERK1/2的磷酸化。SA-B不影响HSCs上TβR I和TβR II的表达。SA-B降低了HSCs中I型胶原的合成；SA-B也降低了TGF-β1刺激下HSCs中I型胶原的合成和分泌。SA-B对MMP-2和MMP-13的活性无影响，但诱导了MMP-13的活性。
SA-B抑制HSC中TGF-β1诱导的ERK信号通路。这种抑制与HSCs上TβR I和TβR II的表达无关。SA-B通过抑制TGF-β1信号通路降低HSC中I型胶原的合成和分泌，这可能与基质金属蛋白酶的降解活性无关。
