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目的:研究大豆异黄酮对血管紧张素Ⅱ(AngⅡ)诱导的血管平滑肌细胞(VSMC)增殖的影响并探讨其作用机制。方法:分离大鼠胸主动脉,组织贴块法培养VSMC,以AngⅡ为诱导剂,建立VSMC细胞增殖模型。应用MTT法,CellTiter-GloR As-say法检测细胞增殖,观察不同浓度的大豆异黄酮及亚硝基-精氨酸甲酯(L-NAME)对AngⅡ诱导的VSMC增殖的影响。硝酸还原酶法检测细胞上清液中一氧化氮(NO)含量;化学比色法测定一氧化氮合酶(NOS),诱导型一氧化氮合酶(iNOS)水平;半定量RT-PCR检测VSMC iNOS mRNA的表达。结果:大豆异黄酮能剂量依赖性的抑制VSMC增殖,该作用可被L-NAME部分抵消;大豆异黄酮能升高NOS、iNOS和NO水平,增加iNOS mRNA的表达。结论:大豆异黄酮可呈剂量依赖性的抑制AngⅡ诱导的VSMC增殖,该作用可能通过上调iNOS基因表达、升高NO水平实现的。
Objective: To study the effect of soybean isoflavones on the proliferation of vascular smooth muscle cells (VSMC) induced by angiotensin II (AngII) and to explore its mechanism of action. METHODS: Rat thoracic aorta was isolated, VSMCs were cultured with tissue patch method, and Ang II was used as inducer to establish VSMC cell proliferation model. Cell proliferation was detected by MTT assay and CellTiter-GloR As-say method. The effects of different concentrations of soybean isoflavones and nitroso-arginine methyl ester (L-NAME) on proliferation of VSMCs induced by AngII were observed. Nitric oxide reductase assay was used to detect the content of nitric oxide (NO) in the supernatant of the cells; chemical colorimetry was used to determine the levels of nitric oxide synthase (NOS) and inducible nitric oxide synthase (iNOS); semi-quantitative RT-PCR detection VSMC iNOS mRNA expression. Results: Soybean isoflavones could inhibit the proliferation of VSMC in a dose-dependent manner. This effect could be offset by L-NAME. Soybean isoflavones could increase the levels of NOS, iNOS and NO and increase the expression of iNOS mRNA. Conclusion: Soybean isoflavone can inhibit the proliferation of VSMC induced by AngII in a dose-dependent manner. This effect may be achieved by up-regulating iNOS gene expression and increasing NO level.