雌激素/雌激素受体α信号通路抑制血管平滑肌细胞SM22α的转录作用

摘要

目的 确定雌激素/雌激素受体α(ERα)信号通路对大鼠血管平滑肌细胞(VSMCs)分化标志基因SM22α的转录调节作用.方法 利用组织块贴壁法分离获得大鼠VSMCs,采用PCR方法扩增获得大鼠SM22α,并插入到pGL3-Basic质粒中构建SM22α启动子荧光素酶报告基因质粒.分别在VSMCs和COS-7细胞中利用Luciferase检测雌二醇(E2)、ERα及Myoeardin对SM22α启动子转录活性的影响情况.结果 SM22α启动子荧光素酶报告基因质粒构建成功,E2、ERα可以抑制Myocardin对SM22α的转录活性.结论 E2/ERα信号通路可通过抑制Myocardin对SM22α的转录活性,从而促进VSMCs由分化型向增殖型转变.%Purpose To confirm the transcriptional regulation effect of estradiol/estrogen receptor a (ERα) pathway on SM22α,the most important differentiation marker in rat vascular smooth muscle cells (VSMCs). Methods Rat VSMCs were isolated by using the method of tissue adherent. The rat SM22a gene was amplified by PCR and inserted into pGL3-Basic vector to construct the SM22α promoter lucifer-ase reporter plasmid. The effect of estradiol ( E2) , ERα and myocardin on the transcription activity of SM22α promoter was detected by the method of luciferase reporter assay in YSMCs and COS-7 respectively. Results SM22α promoter luciferase reporter gene plasmid was constructed successfully. The results of luciferase reporter assay showed that E2 and ERα could inhibit the transactivate effect of myocardin on SM22α. Conclusion E2/ERα signaling pathway can promote the VSMCs to proliferation from differentiated transformation by inhibiting the transcriptional activity of myocardin on SM22α.