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[摘要]
目的 研究川芎嗪与黄芪甲苷配伍对缺氧诱导损伤的人脐静脉内皮细胞(HUVECs)的增殖的影响及机制。方法 建立HUVECs缺氧损伤模型,将细胞分为5组,对照组、模型组、川芎嗪(80 μg/mL)组、黄芪甲苷(40 μg/mL)组及川芎嗪(80 μg/mL)+黄芪甲苷(40 μg/mL)组,MTT法检测川芎嗪、黄芪甲苷及其配伍对缺氧损伤HUVECs增殖的影响;免疫组化法检测缺氧损伤HUVECs细胞VEGF、Ang-II蛋白表达,Western blotting检测VEGF和Ang-II蛋白的表达,RT-PCR检测VEGF和Ang-II mRNA表达。结果 与模型组相比,川芎嗪与黄芪甲苷均能提高细胞存活率,川芎嗪与黄芪甲苷配伍组具有极显著差异(P<0.001);川芎嗪与黄芪甲苷配伍提高VEGF、Ang-II蛋白表达。同时,川芎嗪与黄芪甲苷配伍组能上调VEGF和Ang-II mRNA的表达(P<0.001)。结论 川芎嗪与黄芪甲苷配伍可能通过增加血管生成靶向因子VEGF和Ang-II的表达发挥促进血管生成的作用。
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[Abstract]
Objective To study the protective effect and mechanism of ligustrazine combined with astragaloside IV on hypoxia injury of human umbilical vein endothelial cells (HUVECs). Methods The model of hypoxia injury was established, and the cells for pharmacodynamics study were divided into five groups:control group, model group, ligustrazine group (80 μg/mL), astragaloside IV group (40 μg/mL), and compatibility of astragaloside IV and ligustrazine group. The effects of ligustrazine, astragaloside IV, and their compatibility on cell proliferation in each group after hypoxia injury were detected by MTT assay. Immunohistochemical method was used to observe the expression of proteins VEGF and Ang-II in HUVECs with hypoxia injury, and Western blotting was used to observe the expression of proteins VEGF and Ang-II. RT-PCR was used to observe the mRNA expression of VEGF and Ang-II. Results Compared with those in the model group, cell viability of ligustrazine group, astragaloside IV group, and compatibility group significantly increased, and the best group was ligustrazine (80 μg/mL)+astragaloside IV (40 μg/mL) group. Ligustrazine (80 μg/mL)+astragaloside IV (40 μg/mL) group could up-regulate the protein expression levels of VEGF and Ang-II and the levels of VEGF and Ang-II mRNA. Conclusion Ligustrazine combined with astragaloside IV may be targeted by increasing angiogenesis factor, and the expression of VEGF and Ang-II plays the role of promoting angiogenesis.
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[基金项目]
国家自然科学基金资助项目(81173597);吉林省卫生计生青年科研课题(2014Q046)