脱氢表雄酮对氧化型低密度脂蛋白诱导的平滑肌细胞单核细胞趋化蛋白1表达的影响及其机制

Study on the Influence of Dehydroepiandrosteroneon on the Expression of Monocyte Chemoattractant Protein-1 in Vascular Smooth Muscle Cells Induced by Oxidized Low Density Lipoprotein and Its Mechanism

目的观察脱氢表雄酮对氧化型低密度脂蛋白诱导的血管平滑肌细胞分泌单核细胞趋化蛋白1的影响,并探讨其作用机制是否与细胞色素P450芳香酶的催化作用有关。方法使用脂质体转染法将含有细胞色素P450芳香酶基因的质粒和空白对照质粒分别转染至体外培养的血管平滑肌细胞,24h后给予氧化型低密度脂蛋白诱导及脱氢表雄酮刺激,采用逆转录聚合酶链反应、实时荧光定量聚合酶链反应及酶联免疫吸附法检测转染后各组细胞单核细胞趋化蛋白1的基因和蛋白表达水平。结果与氧化型低密度脂蛋白刺激组比较,给予氧化型低密度脂蛋白和脱氢表雄酮后单核细胞趋化蛋白1的分泌明显降低(P<0.05)。转染含有细胞色素P450芳香酶基因的质粒组和空白对照质粒组单核细胞趋化蛋白1的分泌差别不明显(P>0.05)。结论脱氢表雄酮能抑制氧化型低密度脂蛋白诱导的血管平滑肌细胞单核细胞趋化蛋白1的分泌升高,可能是其抗动脉粥样硬化的机制之一。而这一过程可能并不通过转化为雌雄激素而发挥,也许与其本身的生物学活性有关。

Aim To observe the effects of dehydroepiandrosterone on expression of monocyte chemoattractant protein-1 （MCP-1） induced by oxidized low density lipoprotein （ox-LDL） in vascular smooth muscle cells, and investigate whether its mechanism has something to do with the catalysis of the cytochrome P450 aromatase （CYP19）. Methods Transient transfected the plasmid with or without CYP19 into cultured vascular SMC respectively by lipidosome transfection. 24 h later, the cells were stimulated with ox-LDL and DHEA. Using the method of RT-PCR, Realtime PCR, ELISA, the gene and protein of MCP-1 expression levels of each group were detected. Results Compared with the group stimulated by ox-LDL, the secretion level of MCP-1 was obviously reduced after given ox-LDL and DHEA （P〈0.05）. There wasn’t obvious difference of MCP-1 expression in the groups of transfected plasmid with or without CYP19 （P〉0.05）. Conclusion DHEA shows inhibiting effects on ox-LDL-induced MCP-1 expression in vascular SMC, which may be one of the mechanisms of its antiatherosclerosis. And this effect is not mediated by CYP19.