淫羊藿素对大鼠肝星状细胞HSC-T6的抑制作用  

Suppressive effects of Icaritin on HSC-T6 cells of rats

作　　者：刘进锴[1] 阙任烨 龙建云[1] 李静[1] 黄亮[1] 严以群[1] 

机构地区：[1]第二军医大学附属东方肝胆外科医院肝外一科,上海200433 [2]上海中医药大学附属市中医医院消化内科,上海200071

出　　处：《肝胆胰外科杂志》2014年第4期317-322,共6页Journal of Hepatopancreatobiliary Surgery

摘　　要：目的探讨淫羊藿素对肝星状细胞HSC-T6的影响及其相关机制。方法体外培养大鼠肝星状细胞HSC-T6,用含10%胎牛血清的无酚红DMEM培养基培养细胞,细胞用含5%活性炭吸附胎牛血清的无酚红DMEM培养基接种于细胞培养板。采用雌激素受体完全拮抗剂ICI-182780观察淫羊藿素(ICT)是否能够通过雌激素受体对HSC-T6细胞产生影响。实验分为对照组、ICI-182780组、淫羊藿素组、淫羊藿素联合ICI-182780组。通过MTT和免疫组化来观察ICT对HSC-T6增殖和活化的影响,通过ELISA检测TGF-β1、TIMP-1、MMP-1的表达活性来观察ICT对细胞外基质合成与降解的影响,通过Western blotting的方法检测ERK、p38、JNK的磷酸化水平来观察ICT对MAPK信号通路的影响。结果与对照组比较,加入ICI-182780(1μM)对HSC-T6细胞增殖、a-SMA、TGF-β1、TIMP-1、MMP-1的表达以及pERK、pp38、pJNK的水平均无明显影响。药物ICT干预后,细胞增殖、a-SMA、TGF-β1、TIMP-1的表达以及pERK、pp38、pJNK的水平明显被抑制,MMP-1的表达活性明显增强,差异有统计学意义(P<0.01),药物ICT联合ICI与单独用药比较差异无统计学意义(P>0.05)。结论 ICT可以抑制HSC-T6细胞的增殖与活化、降低细胞外基质的合成,但是我们发现上述效应不是通过雌激素受体来调控的。Objective To investigate the effects of icaritin （ICT） on HSC-T6 cells of rats and the associated mechanisms. Methods Rats HSC-T6 cell line was selected, and cultured in phenol red-free DMEM with 10% FBS. Cells were then cultured in 5% sFBS phenol red-free DMEM. A pure estrogen receptor antagonist was used to observe whether the effects of ICT on HSC-T6 cells were acted through estrogen receptor. The experiment were divided into 4 groups, including control group, ICI-182780 group, icaritin group and ICT＋ICI-182780 group. The proliferation and activation of HSC-T6 cells were detected by MTT assay and immunocytochemistry respectively. TGF- β1, TIMP-1, MMP-1 were examined to observe the effects of ICT on synthesis and degradation of the extracellular matrixc （ECM） by ELISA test. The phosphorylation of MAPK signal pathway was detected by western blot analysis to find out the molecular mechanism of these effects. Results Compared with the control group, the administration of ICI-182780 （1 μmol/L） had no significant effect on any indicators involved in the experiment. Under treatment of ICT, the cell proliferation, as well as alpha-SMA, TGF- β1 and TIMP-1 expres- sion were significantly inhibited （P〈0.01）. On the contrary, MMP-1 expression was increased. The phosphorylation of MAPK signal pathway including ERK, JNK, P38 was obviously decreased （P〈0.01）. However, these effects of ICT could not be blocked by ICI-182780. Conclusion ICT can inhibit HSC-T6 cells proliferation and activation, modulate the synthesis and degradation of ECM. These effects may be associated with the suppression of MAPK signal pathway phosphorylation in a ER-independent way.

关 键 词：淫羊藿素 肝星状细胞HSC-T6 雌激素受体 丝裂原激活蛋白激酶 大鼠 

分 类 号：R285.5[医药卫生—中药学]