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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 2 4、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
OBJECTIVE: To investigate the effects of Ang Ⅱ on the proliferation of vascular smooth muscle cells (VSMCs) and the regulation of human α1 (Ⅱ) chain promoter activity and to understand the role of AngⅡ in the process of atherosclerosis Role. Methods: ① Thoracic aorta VSMCs were cultured in vitro. MTT assay was used to detect the effects of different concentrations of AngⅡ (10 8mol / L, 10 7mol / L, 10 6mol / L) on VSMCs proliferation at 2, 4, . (2) To construct a recombinants containing 2.4 kb and 1.6 kb flanking 5 ’flanking sequences of human α2 (Ⅰ) procollagen gene and chloramphenicol acetyltransferase (CAT) reporter gene, transient transfection of smooth muscle cells with FuGENE transfection reagent, CAT The effect of Ang Ⅱ 10 7mol / L on recombinant was determined by ELISA. Results AngⅡ could promote VSMC proliferation in a dose-dependent manner. AngⅡ group relative CAT expression was significantly increased, with statistical significance. CONCLUSION: AngⅡ can promote the proliferation of VSMC and upregulate the gene expression of α2 (Ⅰ) procollagen at the transcriptional level to promote the formation and development of atherosclerosis