【摘要】 目的研究丹参单体IH764-3对H2O2刺激的肝星状细胞(HSCs)胶原合成、膜型基质金属蛋白酶(MT1-MMP)、基质金属蛋白酶2(MMP-2)及其组织抑制因子(TIMP-2)表达的影响,探讨丹参单体IH764-3在调控HSCs胶原代谢中的作用机制。方法应用体外细胞培养技术,以不同剂量IH764-3干预过氧化氢(H2O2)刺激的HSCs,通过3 H-脯氨酸(3 H-Pro)掺入法检测HSCs总胶原和Ⅰ型胶原合成能力,应用Western blot技术检测MT1-MMP、MMP-2及TIMP-2蛋白的表达。结果不同剂量IH764-3(10μg/ml,20μg/ml,30μg/ml,40μg/ml)作用于H2O2刺激的HSCs 24小时,与单纯H2O2组相比,30μg/ml组和40μg/ml组能显著抑制总胶原和Ⅰ型胶原的合成(P<0.05),而H2O2组、10μg/ml组和20μg/ml组之间差异无统计学意义(P>0.05)。IH764-3干预组与H2O2组相比,能够明显上调MT1-MMP和MMP-2的表达,下调TIMP-2表达,并呈剂量依赖性关系。结论丹参单体IH764-3可能通过下调TIMP-2、上调MT1-MMP及MMP-2表达调控HSCs胶原代谢。更多还原

【Abstract】 Objective To better understand the effects of different concentrations of salvia miltiorrhiza monomer IH764-3 on collagen metabolism in hepatic stellate cells(HSCs),the expression of membrane-type matrix metalloproteinase-1(MT1-MMP),matrix metalloproteinase-2(MMP-2) and tissue inhibitor of metalloproteinase-2(TIMP-2) were investigated.Methods Activated HSCs were cultured in vitro prestimulated by H2O2.After treatment with different concentrations(10,20,30,40 μg/ml) of IH764-3 for 24 hours,the contents of total collagen and type Ⅰ collagen were detected by 3H-proline incorporation assay.The expression of MT1-MMP,MMP-2 and TIMP-2 were determined by western blot.Results The 30 μg/ml group and 40 μg/ml group of IH764-3 markedly inhibited total collagen and type Ⅰ collagen synthesis compared to those of the control group(P<0.05).There were no significant differences among the control group,10 μg/ml group and 20 μg/ml group(P>0.05).The IH764-3 groups(10,20,30,40 μg/ml) could up-regulate the expression of both MT1-MMP and MMP-2 and down-regulate the TIMP-2 in dose-dependent pattern.Conclusion IH764-3 can inhibit the expression of total collagen and type Ⅰ collagen in dose-dependent pattern,the mechanism may be related to the up-regulation of both MT1-MMP and MMP-2 and the down-regulation of TIMP-2.更多还原