摘要
目的探讨miR-26a在血小板源性生长因子B(PDGF-BB)诱导的血管平滑肌细胞(VSMCs)表型转换中的调控作用。方法培养原代小鼠主动脉VSMCs,将细胞分空白对照组、anti-miR-26a、PDGF-BB+anti-miR-control组和PDGF-BB+anti-miR-26a组共4组。分别加入荧光蛋白(Ad-GFP)、anti-miR-26a、PDGF-BB+anti-miR-control和PDGFBB+anti-miR-26a,观察VSMCs的表型改变;用RT-qPCR及Western blot分别检测VSMCs中α平滑肌肌动蛋白(α-SMA)、平滑肌肌球蛋白重链(SM-MHC)、钙调节蛋白(calponin)基因的mRNA和蛋白表达水平以及VSMCs中miR-26a表达。结果与对照组相比,PDGF-BB以浓度和时间依赖方式抑制VSMCs分化标志基因α-SMC、calponin、SM-MHC的mRNA和蛋白的表达(P<0. 05),诱导VSMC表型转换为合成表型; PDGF-BB处理的VSMCs中miR-26a表达显著升高(P<0. 05);抑制miR-26a后,PDGF-BB对VSMCs的α-SMC、calponin、SM-MHC的mRNA和蛋白的抑制作用被部分抵消(P<0. 05)。结论 PDGF-BB使VSMCs中miR-26a表达显著升高,miR-26a促进VSMCs增殖和迁移,miR-26a在PDGF-BB诱导VSMCs表型转换中可能起关键作用。
Objective To investigate the role of miR-26a played in the PDGF-BB induced VSMC phenotypic transformation.Methods Primary mouse aortic VSMCs were divided into blank control group,anti-miR-26a group,PDGF-BB+anti-miR-control group and PDGF-BB+anti-miR-26a group treated with Ad-GFP,anti-miR-26a,PDGF-BB+anti-miR-control and PDGF-BB+anti-miR-26a respectively.Then VSMCs phenotypic transformation was observed.The mRNA and protein expression of alpha-SMA,calponin,SM-MHC and VSMC differentiation marker genes,were detected by RT-PCR and Western blot respectively.The expression of miR-26a was determined by real-time PCR.Results Compared with control group,PDGF-BB induced a dose-dependent and time-dependent suppression of the mRNA and protein levels of α-SMA,calponin and SM-MHC(P<0.05);PDGF-BB significantly increased the miR-26a expression in VSMCs(P<0.05);Inhibition of miR-26a partly abrogated the depression of PDGF-BB on alpha-SMA,calponin and SM-MHC(P<0.05).Conclusions PDGF-BB increases the expression of miR-26a in VSMCs.MiR-26a promotes cell proliferation and migration of VSMCs,miR-26a may play a critical role in PDGF-BB induced VSMCs phenotype transformation.
作者
洪新宝
王炳才
余江水
杨晓燕
郭榕
顾信建
李月婷
HONG Xin-bao;WANG Bing-cai;YU Jiang-shui;YANG Xiao-yan;GUO Rong;GU Xin-jian;LI Yue-ting(Department of Cardiology,the Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China;Department of Nephrology,the Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China)
出处
《基础医学与临床》
CSCD
2019年第2期192-196,共5页
Basic and Clinical Medicine
基金
福建省自然科学基因引导性项目(2016Y0032)
