加味茵陈四逆汤含药血清影响肝星状细胞胶原蛋白表达的机制研究

Effect of Drug Serum of Modified Yinchen Sini Decoction on Proliferation of Rat Hepatic Stellate Cells

目的:探讨加味茵陈四逆汤对大鼠肝星状细胞Collagen1及MMP-1/TIMP-1的影响。方法:以HSC-T6细胞株为主要研究对象,将其依据实验目的分为6组,分别为空白组,模型组,阳性药物组,中药高、中、低剂量组。除空白组无需特殊处理外,其余5组给予含药血清培养同时给予细胞因子TGF-β1促进细胞增殖,培养48h后收集细胞,提取蛋白,并采用Western blot方法检测各观察组细胞Collagen1及MMP-1/TIMP-1的表达,并比较其差异。结果:与空白组比较,模型组MMP-1/TIMP-1、Collagen1蛋白表达均升高(P<0.05);与模型组比较,加味茵陈四逆汤中、低剂量组MMP-1蛋白表达升高(P<0.05),而Collagen1在中药中、低剂量组表达明显降低(P<0.05),TIMP-1在中药低剂量组明显降低(P<0.05)。结论:加味茵陈四逆汤含药血清可能通过提高MMP-1的表达以及降低TIMP-1从而降低胶原蛋白表达,最终延缓肝纤维化的发生。

Objective:To investigate the effect of drug serum of modified Yinchen Sini Decoction on MMP-1/TIMP-1 and Collagen 1 of TGF-β1-induced rat hepatic stellate cells(HSCs).Methods:Rat hepatic stellate cells were the study objects,which were divided into six groups:blank group,model group,positive drug group,high,middle and low dose groups of Chinese medicine.All groups were given TGF-β1 except blank group and cultured with 10% drug serum culture.The protein expression levels of MMP-1/TIMP-1 and Collagen 1 in HSC-T6 cells of all groups were detected by using Western blot assay.Results:As compared with the blank group,protein expression levels of MMP-1/TIMP-1 and Collagen 1 were increased(P<0.05).As compared with the model group,the protein expression level of MMP-1 was significantly increased in modified Yinchen Sini Decoction middle and low dose groups(P<0.05),however Collagen 1 was significantly decreased(P<0.05).The protein expression levels of TIMP-1 were decreased in modified Yinchen Sini Decoction low dose group(P<0.05).Conclusion:Drug serum of modified Yinchen Sini Decoction could up-regulate the protein expression levels of MMP-1 and down-regulate TIMP-1,suggesting that the drug can delay the process of liver fibrosis under the in vitro conditions.