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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。
AIM: To investigate the effect and mechanism of Retinol-binding protein 4 (RBP4) on the migration and proliferation of vascular smooth muscle cells (SMCs). Methods: The aortic SMCs were cultured in vitro and the effects of RBP4 on the migration of SMCs were observed by scratch test and Boyden’s migration chamber assay. The phosphorylation of Akt was detected by Western blotting. The PI3K inhibitor LY294002 pretreatment Cells on RBP4-induced migration of SMCs, the application of MTT colorimetric assay and flow cytometry techniques to detect RBP4 SMCs proliferation and cell cycle. RBP4-treated cells significantly increased Akt phosphorylation; PI3K inhibitor LY294002 pre-treated cells significantly inhibited RBP4-induced migration (P <0.05) RBP4 treatment had the tendency to increase the number of SMCs, and could slightly block the cells entering the S phase, but did not reach statistical significance (P> 0.05). CONCLUSION: RBP4 induces the migration of vascular SMCs via PI3K-Akt pathway, but has no significant effect on cell proliferation and cell cycle.