Shanghai Shuguang Hospital, PR China.
J Ethnopharmacol. 2010 Nov 11;132(2):384-92. doi: 10.1016/j.jep.2010.05.026. Epub 2010 Jul 3.
Salvianolic-acid B (SA-B) is an effective component of Radix Salviae miltiorrhizae for anti-hepatic fibrotic herbs. MAPK signaling pathway has been implicated in hepatic stellate cells (HSC) stimulated by TGF-(1. We have investigated the effect of SA-B on MAPK pathway in rat HSC.
To observe the pharmacological effect of SA-B on HSC, SA-B was added into the medium of primary HSC. TGF-(1 was added during last 2h, and PD98059 (ERK inhibitor) and SB203580 (p38 inhibitor) were added just 30 min before adding TGF-(1. MEF2 and Col. I were measured by luciferase reporter gene assay and Western blot. (-SMA, MEF2, Raf, ERK, p-ERK, MEK, p-MEK, p38, p-p38, MKK3 and p-MKK3/6 were assayed by Western blot. Activity of MMP-2 and MMP-9 was analyzed by zymography. Each experiment was repeated for three times.
The expression of (-SMA and Col. I in HSC was inhibited by SA-B. There was no effect of SA-B on the activity of MMP-2 or MMP-9 in the media of cultured HSC. Phosphorylation of ERK1/2 in HSC stimulated with or without TGF-(1 was inhibited by SA-B. Specifically, phosphorylation of MEK (upstream kinase of ERK pathway) was inhibited by SA-B. SA-B also inhibited phosphorylation of MKK3/6 (upstream kinases of p38 pathway) and inhibited the synthesis of MEF2.
SA-B performs anti-hepatic fibrosis through inhibiting ERK and p38 MAPK pathway in HSC. SA-B inhibits ERK pathway via inhibiting phosphorylation of MEK and inhibits p38 MAPK pathway via blocking phosphorylation of MKK3/6 and inhibiting expression of MEF2 in HSC with or without TGF-(1 stimulation.
丹酚酸 B（SA-B）是丹参抗肝纤维化的有效成分之一。MAPK 信号通路已被证实参与了转化生长因子-β1（TGF-β1）刺激的肝星状细胞（HSC）。本研究旨在探讨 SA-B 对大鼠 HSC 中 MAPK 通路的影响。
为了观察 SA-B 对 HSC 的药理作用，将 SA-B 加入原代 HSC 的培养基中。在加入 TGF-β1 的前 2 小时加入 SA-B，并在加入 TGF-β1 前 30 分钟加入 PD98059（ERK 抑制剂）和 SB203580（p38 抑制剂）。通过荧光素酶报告基因检测和 Western blot 检测 MEF2 和 Col. I 的表达。通过 Western blot 检测 (-SMA、MEF2、Raf、ERK、p-ERK、MEK、p-MEK、p38、p-p38、MKK3 和 p-MKK3/6 的表达。通过明胶酶谱分析 MMP-2 和 MMP-9 的活性。每个实验重复三次。
SA-B 抑制 HSC 中 (-SMA 和 Col. I 的表达。SA-B 对培养的 HSC 培养基中 MMP-2 或 MMP-9 的活性没有影响。SA-B 抑制 TGF-β1 刺激或不刺激的 HSC 中 ERK1/2 的磷酸化。具体来说，SA-B 抑制 MEK（ERK 通路的上游激酶）的磷酸化。SA-B 还抑制 MKK3/6（p38 通路的上游激酶）的磷酸化，并抑制 MEF2 的合成。
SA-B 通过抑制 HSC 中的 ERK 和 p38 MAPK 通路发挥抗肝纤维化作用。SA-B 通过抑制 MEK 的磷酸化抑制 ERK 通路，通过阻断 MKK3/6 的磷酸化和抑制 TGF-β1 刺激或不刺激的 HSC 中 MEF2 的表达抑制 p38 MAPK 通路。
