复方玉郎伞对肝星状细胞的影响

Effect of compound Yu Lang San in HSC-T_6 cells

目的:研究复方玉郎伞(CYLS)对肝星状细胞(HSC-T6)增殖、细胞上清液中Hyp含量及基质金属蛋白酶-2(MMP-2)、金属蛋白酶组织抑制因子-2(TIMP-2)mRNA表达的影响。方法:体外培养HSC-T6,四甲基偶氮唑盐比色(MTT)法检测玉郎伞多糖(YLSPS)、玉郎伞黄酮(YLSYF)及六月青多糖(LYQPS)对HSC-T6的抑制作用,以细胞存活率为90%的各药物浓度配制成为最大浓度CYLS应用液,再次以MTT法测定其对HSC-T6的抑制作用。以细胞存活率大于90%的各药物浓度配制成3种不同配比的CYLS应用液。以CYLS应用液对HSC-T6进行干预,消化法检测细胞上清液中羟脯氨酸(Hyp)含量,逆转录聚合酶链反应(RT-PCR)法测定其对MMP-2,TIMP-2 mRNA的影响。结果:CYLS各配方均可不同程度地降低细胞上清液中Hyp的含量;配方一和配方二能显著上调HSC-T6中MMP-2 mRNA的表达;各配方均能显著下调HSC-T6中TIMP-2 mRNA表达。结论:CYLS可抑制胶原蛋白的生成、促进细胞外基质(ECM)的降解,这可能是CYLS抗肝纤维化的作用机制之一。

Objective：To study the effect of compound Yu Lang San （CYLS） on proliferation,content of Hyp in the supernatant of cells,and the mRNA expressions of MMP-2 and TIMP-2 in HSC-T6 cells. Methods：HSC-T6 cells were cultured in vitro. MTT assay was used to evaluate the inhibitive effect of YLSPS,YLSYF and LYQPS in HSC-T6 cells. The concentrations of YLSPS,YLSYF and LYQPS for 90% survival of HSC-T6 cells were chosen to prepare cocktail mixtures of the highest concentrations as evaluated by MTT assay. The concentrations causing above 90% HSC-T6 cell survival were chosen to mix into 2 kinds of CYLS. After HSC-T6 cells were treated with CYLS,the content of Hyp in the supernatant of cells was detected by digestion assay,and MMP-2 and TIMP-2 mRNAs were determined by RT-PCR. Results：Three kinds of CYLS reduced the content of Hyp in the supernatant of cells and down-regulated the expression of TIMP-2 mRNA; the compounds 1 and 2 up-regulated the expression of MMP-2. Conclusion：CYLS can reduce ECM synthesis and promote ECM degradation,which may be one of the possible molecular mechanisms of inhibiting hepatic fibrosis.