PDGF-BB对血管平滑肌细胞表型标志物表达的影响  

Effect of PDGF-BB on expression of phenotypic markers in vascular smooth muscle cells

作　　者：高蕊[1] 董丽华[1] 谢肖立[1] 方新梅[1] 温进坤[1] 韩梅[1] 

机构地区：[1]河北医科大学基础医学研究所生物化学与分子生物学研究室神经与血管生物学省部共建教育部重点实验室,河北石家庄050017

出　　处：《中国病理生理杂志》2010年第12期2301-2305,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31071003);河北省自然科学基金资助项目(No.C2007000831)

摘　　要：目的:观察血小板源性生长因子BB(PDGF-BB)对血管平滑肌细胞(VSMCs)增殖及分化相关基因表达的影响,探讨其可能的机制。方法:分离体外培养的SD大鼠胸腹主动脉VSMCs,分为空白对照组和不同浓度PDGF-BB处理组。分别采用MTT法、流式细胞术和伤口愈合实验检测PDGF-BB对VSMCs增殖、细胞周期和迁移活性的影响;用W estern b lotting分析检测VSMCs表型标志物的表达;用免疫沉淀和免疫共沉淀分析检测Krüppel样因子4(KLF4)磷酸化及与其它转录因子的相互作用。结果:PDGF-BB促进VSMCs增殖和迁移;上调增殖相关蛋白PCNA的表达,下调增殖抑制蛋白p27、分化相关蛋白SM22α的表达。PDGF-BB诱导KLF4的表达和磷酸化,促进KLF4与NF-κB的相互作用,抑制KLF4与Sm ad3、HDAC2的结合。结论:PDGF-BB可能通过影响KLF4磷酸化及其与不同转录调节因子的相互作用而诱导VSMCs表型转化。AIM：To study the effect of platelet-derived growth factor（PDGF）-BB on the expression of phenotypic markers in vascular smooth muscle cells（VSMCs） and to investigate the underlying mechanisms in phenotypic switching and proliferation of VSMCs.METHODS： Cultured rat VSMCs were treated with PDGF-BB.The cell viability and cell cycle were detected by MTT assay and flow cytometry,respectively.The cell migration was examined by wound-healing assay.The protein expression was examined by Western blotting.Immunoprecipitation and co-immunoprecipitation were used to detect KLF4 phosphorylation and its interaction with other transcription factors,respectively.RESULTS： PDGF-BB induced the proliferation and migration of VSMCs.PDGF-BB increased the expression of PCNA,KLF4 and phosphorylated KLF4,and reduced the levels of p27 and SM22α.Moreover,the interaction of KLF4 with NF-κB was increased,and the interaction of KLF4 with Smad3 or HDAC2 was decreased by the action of PDGF-BB.CONCLUSION： KLF4 plays a role in the proliferation of VSMCs induced by PDGF-BB,which is related with its phosphorylation and the interaction with other transcription factors.

关 键 词：血管平滑肌细胞 血小板源性生长因子 Krüppel样因子4 表型 

分 类 号：R34[医药卫生—基础医学]