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目的:观察中药复方丹参对肝星状细胞胶原合成与降解的影响。方法:选择肝星状细胞系作为体外研究对象,MTT法观察药物对细胞增殖的影响,ELISA法检测细胞上清液Ⅰ、Ⅲ、Ⅳ型胶原含量,RNA-cDNA分子杂交检测细胞内Ⅰ、Ⅲ、Ⅳ型胶原及TGF β1 mRNA稳态水平。逆转录多聚酶链反应检测间质胶原酶及金属蛋白酶组织抑制因子-1基因表达水平。结果:中药复方丹参明显抑制肝星状细胞增殖且呈剂量依赖趋势,与对照组比较,中药组细胞上清液中Ⅰ、Ⅲ、Ⅳ型胶原含量明显降低,细胞内Ⅰ、Ⅲ、Ⅳ型胶原及TGF β1 mRNA稳态水平明显降低,间质胶原酶基因表达水平明显增强,金属蛋白酶组织抑制因子-1基因表达水平明显受到抑制(P<0.01)。结论:中药复方丹参可能通过抑制肝星状细胞的增殖,抑制胶原的异常合成,促进异常沉积胶原的降解,从而起到抗肝纤维化的作用。
OBJECTIVE: To observe the effect of Chinese compound salvia miltiorrhiza on the synthesis and degradation of collagen in hepatic stellate cells. METHODS: Hepatic stellate cell line was selected as in vitro study. MTT assay was used to observe the effects of drugs on cell proliferation. ELISA was used to detect collagen I, III, and IV in supernatants. RNA-cDNA hybridization was used to detect intracellular I and III. , type IV collagen and TGFβ1 mRNA level. Retrograde transcription polymerase chain reaction was used to detect the expression of interstitial collagenase and tissue inhibitor of metalloproteinase-1 gene. RESULTS: Chinese herbal compound Salvia miltiorrhiza can obviously inhibit the proliferation of hepatic stellate cells in a dose-dependent manner. Compared with the control group, the content of type I, III, IV collagen in the supernatant of the traditional Chinese medicine group was significantly reduced, and intracellular collagen types I, III, and IV were decreased. And the steady-state level of TGFβ1 mRNA was significantly decreased, the expression level of interstitial collagenase gene was significantly increased, and the expression level of tissue inhibitor of metalloproteinase-1 gene was significantly inhibited (P<0.01). Conclusion: Salvia miltiorrhiza can inhibit the proliferation of hepatic stellate cells, inhibit the abnormal synthesis of collagen, promote the degradation of abnormal deposition collagen, and thus play an anti-hepatic fibrosis effect.