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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 effects of CYLS on the proliferation of hepatic stellate cells (HSC-T6), the level of Hyp in the supernatants and the expression of matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 -2) mRNA expression. Methods: The effects of YLSPS, YLSYF and LYQPS on HSC-T6 in vitro were detected by HSC-T6 and MTT assay. Inhibition, the cell survival rate of 90% of the drug concentration formulated as the maximum concentration of CYLS application solution, again by MTT assay to determine its inhibitory effect on HSC-T6. To the survival rate of more than 90% of the drug concentration formulated into three different proportions of CYLS application solution. HSC-T6 was intervened by CYLS solution, the content of hydroxyproline (Hyp) in the cell supernatant was determined by digestion method, and the effect of MMP-2, TIMP-2 Effect of mRNA. Results: All the formulations of CYLS could reduce the content of Hyp in the supernatant of the cells to a certain extent. Formulation 1 and 2 could significantly up-regulate the expression of MMP-2 mRNA in HSC-T6. All the formulations could significantly down-regulate the expression of TIMP in HSC-T6 -2 mRNA expression. Conclusion: CYLS can inhibit the formation of collagen and promote the degradation of extracellular matrix (ECM), which may be one of the mechanisms of anti-liver fibrosis of CYLS.