肝素对大鼠血管平滑肌细胞同型半胱氨酸诱导的基质金属蛋白酶-2表达的影响

Effects of Heparin on the Production of Homocysteine-induced Extracellular Matrix Metalloproteinase-2 in Cultured Rat Vascular Smooth Muscle Cells

目的研究肝素对大鼠血管平滑肌细胞同型半胱氨酸诱导的基质金属蛋白酶一2（MMP-2）表达的影响。方法利用免疫印迹法和明胶酶谱分析法研究大鼠血管平滑肌细胞中不同浓度的同型半胱氨酸（0～1000umol／L）及不同浓度的肝素（0～1D0ug／m1）对MMP-2表达的影响。结果同型半胱氨酸的浓度在50umol／L、100umol／L、500umol／L、1000umol／L时，MMP-2分别为1．32±012、1．65±0．17、2．48士0．21、2．71±0．24，MMP-2的表达可随同型半胱氨酸浓度的增高而显著增加；作用72hMMP-2的表达与24h和48h比，差异有显著统计学意义（P〈0．01）。在细胞外加入肝素后同型半胱氨酸对MMP-2表达的诱导增强作用可被抑制，当肝素浓度为1．0umol／L、10．Oumol／L、50．0umol／L、100．0umol／L时，MMP-2浓度分别为2．17±0．19、1．67±0．14、1．34±0．14、0．65±0．04。结论大鼠血管平滑肌中同型半胱氨酸可增加MMP-2的表达。细胞外加入肝素可抑制此作用。高同型半胱氨酸血症与动脉粥样硬化发生相关；肝素可防止动脉粥样硬化的发生，对血管具有保护作用。

Objective To study the effects of heparin on the production of homocysteine-induced extracellular matrix metalloproteinase-2 in cultured rats vascular smooth muscle cells. Methods We examined the effects of different homocysteine levels （0 - 1 000 μmol / L） on matrix metalloproteinase-2 （MMP-2） production, and the effects of different heparin concentrations （0 - 100 μg / ml） on homocysteine-induced MMP-2 in cultured rat vascular smooth muscle cells （VSMCs） using gelatin zymography and western blotting. We further compared the changes of MMP-2 under various treatments for 24h, 48h and 72h. Results Homocysteine （50- 1 000 μmol / L） increased the production of MMP-2 significantly in a dose-dependent manner. Increased production of MMP-2 induced by homocysteine was reduced by extracellularly added heparin in a dose-dependent manner. Production of MMP-2 under various treatments for 72h increased more than 24h and 48h. Conclusion Extracellularly added heparin decreased homocysteine-induced MMP-2 secretion. Data suggested a mechanism how hyperhomocysteinemia involved in the pathogenesis of coronary artery disease, and a beneficial effects of heparin on these conditions.