摘要
目的研究白杨素对血小板源性生长因子(PDGF)-BB诱导的血管平滑肌细胞(VSMCs)增殖的影响及细胞周期调控机制。方法WST-1法测定VSMCs增殖,5-溴-2'-脱氧尿苷掺入法测定DNA合成流式细胞术分析VSMCs细胞周期Western blot法测定细胞周期关键调控因子周期蛋白依赖性激酶(CDK)4、CDK6和CDK抑制物p27^(kip1)的表达。结果20ng/ml PDGF-BB显著增强VSMCs增殖及DNA合成,而白杨素呈剂量依赖性抑制PDGF-BB刺激的VSMCs增殖及DNA合成。12.5μmol/L白杨素预处理使PDGF-BB刺激的VSMCs阻滞于G_0/G_1期并能显著降低VSMCs中CDK4和CDK6的蛋白表达水平,同时升高p27^(kip1)蛋白水平。结论白杨素抑制PDGF-BB刺激的VSMCs增殖,其机制可能与CDK4和CDK6表达下调而p27^(kip1)表达上调导致的细胞周期阻滞有关。
Objective To investigate the effects of chrysin on the proliferation of vascular smooth muscle cells （VSMCs） induced by platelet derived growth factor （PDGF）-BB and the possible mechanism. Methods Proliferation of VSMCs was measured by WST-1 assay. DNA synthesis of VSMCs was determined by 5-bromo-2＇-deoxyuridine（BrdU） incorporation. Flow cytometric analysis was carried out to analyze cell cycle of VSMCs. Western blot was performed to evaluate protein levels of cyclin-dependent kinase（CDK）4, CDK6 and p27kip1（a CDK inhibitor）. Results Chrysin inhibited PDGF-BB-induced VSMCs proliferation and DNA synthesis in a concentration-dependent manner. 12.5μmol/L chrysin pretreatment caused PDGF-BB-stimulated VSMCs to arrest at G0/G1 phases, and this arrest was accompanied by decreases in CDK4 and CDK6 protein levels, and an increase in p27kip1expression. Conclusion Chrysin can suppress the proliferation of VSMCs induced by PDGF-BB. The possible mechanism is related to cell cycle arrest mediated by reduced protein levels of CDK4 and CDK6 as well as elevated p27kip1 levels.
出处
《医学研究杂志》
2015年第12期34-37,共4页
Journal of Medical Research
基金
国家自然科学基金资助项目(81000095
30800278)
高等学校新教师基金资助项目(200804861048)
关键词
白杨素
血小板源性生长因子
血管平滑肌细胞
增殖
细胞周期
Chrysin
Platelet derived growth factor
Vascular smooth muscle cells
Proliferation
Cell cycle
