同型半胱氨酸对人主动脉平滑肌细胞ATP结合盒转运蛋白A1启动子甲基化水平及表达的影响
王慧媛+张紫荆+袁封艳[摘要] 目的 探討同型半胱氨酸(Hcy)对人主动脉平滑肌细胞ATP结合盒转运蛋白A1(ABCA1)启动子甲基化水平及表达的影响。 方法 培养人主动脉平滑肌细胞,将其分为两组:对照组和实验组。对照组不加Hcy,实验组加入不同浓度Hcy(50、100、200、500 μmol/L)培养48 h。闪烁计数法检测胆固醇流出率;实时定量PCR检测ABCA1 mRNA表达水平;Western-blotting检测ABCA1蛋白质表达;巢式甲基化PCR检测ABCA1启动子甲基化水平。 结果 对照组各时间点胆固醇流出率比较差异无统计学意义(P > 0.05);各Hcy浓度组不同时间比较显示,24、48 h胆固醇流出率均明显低于0、12 h,差异均有统计学意义(P 0.05),24、48 h各Hcy浓度组胆固醇流出率显著降低,差异均有统计学意义(P 0.05)。 结论 Hcy使人主动脉平滑肌细胞ABCA1启动子甲基化水平升高及表达下降,引起胆固醇流出率降低,可能是其引起动脉粥样硬化的病理机制之一。
[关键词] 同型半胱氨酸;ATP结合盒转运蛋白A1;甲基化;血管平滑肌细胞
[中图分类号] R543.5 [文献标识码] A [文章编号] 1673-7210(2018)01(b)-0019-04
[Abstract] Objective To investigate the effect of homocysteine (Hcy) on the promoter methylation and expression of ATP binding cassette transporter A1 (ABCA1) in human aortic smooth muscle cells. Methods The human aortic smooth muscle cells were divided into two groups: the control group and the experimental group. The cells in the control group were cultured without Hcy, however cells in the experimental group were cultured with different concentrations of Hcy (50, 100, 200, 500 μmol/L). The cholesterol outflow rate was detected by scintillation counting technique ......
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