【摘要】 目的:研究视黄醇结合蛋白4(Retinol-binding protein 4,RBP4)对血管平滑肌细胞(SMCs)迁移和增殖的影响及分子机制。方法:体外培养大鼠主动脉SMCs,采用划痕实验及Boyden’s迁移小室实验观察RBP4对SMCs迁移的影响,采用免疫印迹实验技术检测Akt的磷酸化水平,采用Boyden’s小室实验观察PI3K抑制剂LY294002预处理细胞对RBP4促SMCs迁移的影响,应用MTT比色实验结合流式细胞仪技术,检测RBP4对SMCs细胞增殖及细胞周期的影响。结果:RBP4呈剂量依赖性诱导大鼠血管SMCs迁移(P<0.05);RBP4处理细胞显著增加了Akt磷酸化;PI3K抑制剂LY294002预处理细胞则显著抑制了RBP4的促迁移作用(P<0.05);RBP4处理有增加SMCs数量的趋势,且可轻微阻滞细胞进入S期,但未达到统计学显著性(P>0.05)。结论:RBP4通过PI3K-Akt通路诱导大鼠血管SMCs迁移,对细胞增殖及细胞周期则无显著影响。更多还原

【Abstract】 Objective: To explore a potential role of retinol-binding protein 4(RBP4) in vascular smooth muscle cells(SMCs) migration and proliferation and its molecular mechanism. Methods: Vascular SMCs of rat were cultured in vitro. Vascular SMCs migration was studied by scratch assay and Boyden’s chamber. Western blot analysis was used for detecting Akt and phospho-Akt protein levels.SMCs were pretreated with PI3 K inhibitor LY294002, Boyden’s chamber was then used for detecting RBP4-induced cell migration. Cell proliferation and cell cycle of vascular SMCs induced by RBP4 was detected by use of MTT dye absorbance and flow cytometry. Results:Recombinant human RBP4 significantly induced migration of rat vascular SMCs in a dose-dependent fashion(P<0.05) and phosphorylation of Akt at the same time. PI3 K inhibitor Ly294002 significantly suppressed RBP4’s effects on cell migration(P<0.05). RBP4 treatment slightly increased SMCs proliferation and led to G1/S cell cycle arrest, but these effects did not reach statistical significance. Conclusions: RBP4 induces migration of vascular SMCs through PI3K-Akt-dependent pathways. However, the proliferation and cell cycle progression of the SMCs is not affected by RBP4 treatment.更多还原