同型半胱氨酸对人脐静脉内皮细胞MCP-1 mRNA表达的影响

The Effect of Homocysteine on Monocyte Chemoattractant Protein-1 mRNA Expression in Human Umbilical Vein Endothelial Cells

目的:探讨同型半胱氨酸(Hcy)对人脐静脉内皮细胞单核细胞趋化蛋白-1(MCP-1)mRNA表达的影响。方法:人脐静脉内皮细胞CRL-1730融合至90%分别加入不同浓度Hcy:空白组0mmol/L、生理剂量组0.01mmol/L、病理剂量组0.1mmol/L、非毒性药理剂量组1.0mmol/L。毒性药理剂量组5.0mmol/L,分别刺激细胞6h,用1.0mmol/LHcy刺激内皮细胞1、6、24h;收集细胞提取总RNA,逆转录聚合酶链反应(RT-PCR)测定Hcy对MCP-1mRNA的表达情况。结果:不同浓度Hcy刺激CRL-17306h,细胞内MCP-1mRNA表达差异有统计学意义(P<0.05)。与生理剂量组相比,非毒性药理剂量组MCP-1mRNA表达升高,毒性药理剂量组表达降低;1.0mmol/LHcy刺激CRL-1730不同时间后,MCP-1mRNA表达随刺激时间延长呈逐渐降低的趋势,24h表达量最低(P<0.01)。结论:Hcy可以影响人脐静脉内皮细胞MCP-1mRNA的表达,在动脉粥样硬化的发生过程中发挥重要的作用。

Objective： To investigate the effect of homocysteine （Hey） on the transcription of monocyte chemoattraetant protein-1 （MCP-1）mRNA in human umbilical vein endothelial cells （HUVEC, CRL-1730）. Methods： The HUVEC CRL-1730 were cultured with 1.0 mmol/L Hey at different concentrations （physiological concentration 0.01 retool/L, pathophysiologieal concentration 0.1 mmol/L, pharmacological noncytotoxic concentration 1.0 mmol/L and pharmacological cytotoxic concentration 5.0 retool/L） for 6 h, and with 1.0 mmol/L Hcy in different times （1 h, 6h and 24 h）. The cells were collected to isolate the total RNA, and then RT-PCR was performed. Results： There were significant differences in expression of MCP-1 mRNA in CRL-1730 treated with gradually increased concentrations of Hcy （P 〈 0.05）. The expression of MCP-I mRNA induced by 1.0 mmol/L Hey was down regulated in a time-dependent fashion. Conclusion： The expression of MCP-1 mRNA was affected by homocysteine. The results suggested that homocysteine may contribute to the initiation and progression of atherosclerosis by upregulating MCP-1 expression and promoting leukocyte recruitment.