血管紧张素Ⅱ对血管平滑肌细胞增殖及人I型胶原α2(I)基因启动子活性的影响

The effects of AngⅡon proliferation of vascular smooth muscles and the promoter activities of human α2(Ⅰ) procollagen gene

  • 摘要: 目的: 通过研究血管紧张素Ⅱ(AngⅡ)对血管平滑肌细胞(VSMC)增殖的影响及对人Ⅰ型胶原α2(Ⅰ)链启动子活性的调控,了解AngⅡ在动脉粥样硬化形成和发展过程中的作用。方法: ①SD大鼠胸主动脉VSMC体外培养,分别在24、4 8和72h采用MTT法检测不同浓度的AngⅡ(10-8mol/L、10-7mol/L、10-6mol/L)对VSMCs增殖的影响。②构建含人α2(Ⅰ)前胶原基因5’侧翼序列2.4kb和1.6kb与报告基因氯霉素乙酰基转移酶(CAT)的重组体,用FuGENE转染试剂瞬时转染平滑肌细胞,CAT-ELISA测定AngⅡ 10-7mol/L对重组体的作用。结果: ①AngⅡ剂量依赖性促进VSMC增殖。②AngⅡ组的相对CAT表达量明显增高,有统计学意义。结论: AngⅡ可促进VSMC增殖,并可以在转录水平上调人α2(Ⅰ)前胶原基因表达,从而促进动脉粥样硬化的形成和发展。

     

    Abstract: Objective: To understand the role of AngⅡin the formation and progression of atherosclerosis, the effects of AngⅡon proliferation of VSMC and the modulation on the promoter activities of human α2(Ⅰ) procollagen gene were studied. Methods: 1.Smooth muscle cells derived from SD rat were primary cultured and subcultured. Different concentrations of AngⅡ(10-8 mol/L、10-7 mol/L、10-6 mol/L)were added into the plates to co-incubated with the cells. MTT method was used to detect the cell proliferation after 24, 48 and 72 hours. 2.Cells were transient transfected with pCOLH 22.4 and pCOLH 21.6 containing –2.4kb and –1.6kb of 5’flank sequence of human α2(Ⅰ) procollagen gene, fused to CAT reporter gene by FuGENE Transfection Reagent. The effects of AngⅡ10 -7 mol/L on the plasmid were determined by CAT-ELISA. Results: 1. AngⅡcould accelerate the proliferation of VSMC with dose-dependent. 2. AngⅡ can enhance the promoter activities of human α2(Ⅰ) procollagen gene. Conclusion: These data indicate that AngⅡcan accelerate the proliferation of VSMC and enhance the expression of human α2(Ⅰ) procollagen gene and accelerate the formation and progression of atherosclerosis.

     

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