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本文旨在研究全反式维甲酸(all-trans retinoic acid,ATRA)在血管平滑肌细胞(vascular smooth muscle cell,VSMC)中对apelin基因表达的影响及分子机制。我们用RT-PCR、实时定量PCR和免疫印迹分析检测ATRA对VSMC中apelin基因表达的影响,然后在VSMC中用小干扰RNA转染下调内源性维甲酸受体α(retinoic acid receptorα,RARα)或用腺病毒载体过表达RARα后,检测ATRA对apelin基因表达的影响。结果显示ATRA能以时间和浓度依赖的方式诱导apelin基因的表达,同时RARα表达水平也显著升高,但RARβ和RARγ表达水平无显著变化。利用小干扰RNA下调内源性RARα或用RARα选择性抑制剂Ro 41-5253抑制RARα活性后,再用ATRA刺激VSMC,ATRA对apelin基因表达的诱导作用受到显著抑制,而过表达RARα,则可促进apelin的表达升高。以上结果表明,ATRA可以上调VSMC中apelin基因表达水平,其分子机制是通过其核受体RARα介导完成的。
The purpose of this study was to investigate the effect of all-trans retinoic acid (aTRA) on apelin gene expression in vascular smooth muscle cells (VSMCs) and its molecular mechanism. We detected the effect of ATRA on apelin gene expression in VSMCs by RT-PCR, real-time quantitative PCR and Western blot analysis. Then transfected VSMCs with small interfering RNAs to downregulate endogenous retinoic acid receptorα (RARα) Or adenovirus vector overexpression of RARα, ATRA on apelin gene expression. The results showed that ATRA induced apelin gene expression in a time-and-concentration-dependent manner, meanwhile, the expression of RARα also increased significantly, but the expression of RARβ and RARγ did not change significantly. After down-regulating endogenous RARα with small interfering RNA or inhibiting RARα activity with RARα selective inhibitor Ro 41-5253, ATRA can significantly inhibit apelin gene expression after VSMC stimulation with ATRA, while overexpression of RARα Promote apelin expression increased. The above results show that ATRA can upregulate apelin gene expression in VSMC, and its molecular mechanism is mediated by its nuclear receptor RARα.