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目的:通过缺氧诱导与ox-LDL的相互作用,分析体外缺氧条件下实验兔VSMC增殖、迁移相关基因的表达及生物学行为的改变;探讨缺氧参与VSMC增殖、迁移,加重AS硬化程度的可能机制。方法:原代培养兔主动脉VSMC,传至第3代进行实验。用不同浓度的ox-LDL诱导细胞,并采用低氧环境(<5%O2)进行实验。采用real time PCR和Western Blotting检测HIF1-α、VEGF及其受体、MMP9基因的表达变化;采用BrdU掺入法分析细胞增殖能力;细胞划痕实验检测VSMC的随机迁移能力。
OBJECTIVE: To investigate the effects of hypoxia on the proliferation and migration of VSMCs induced by hypoxia and the changes of biological behavior in VSMCs exposed to hypoxia. To investigate the effects of hypoxia on the proliferation and migration of VSMCs and the severity of AS Possible mechanism. Methods: Primary cultured rabbit aorta VSMC, passed to the third generation of experiments. Cells were induced with different concentrations of ox-LDL and experiments were performed in a hypoxic environment (<5% O2). The expression of HIF1-α, VEGF and its receptor, MMP9 gene were detected by real time PCR and Western Blotting. BrdU incorporation assay was used to analyze cell proliferation ability. Scratch assay was used to detect the random migration ability of VSMC.