miR-26a在血小板源性生长因子B诱导血管平滑肌细胞表型转换的作用

基础医学与临床 ›› 2019, Vol. 39 ›› Issue (2): 192-196.

miR-26a在血小板源性生长因子B诱导血管平滑肌细胞表型转换的作用

洪新宝,王炳才,余江水,杨晓燕,郭榕,顾信建,李月婷

福建医科大学附属第二医院

收稿日期:2018-07-23 修回日期:2018-12-03 出版日期:2019-02-05 发布日期:2019-01-16
通讯作者: 李月婷 E-mail:liyt163@126.com
基金资助:
福建省自然科学基因引导性项目

Effect of miR-26a on phenotypic transformation of vascular smooth muscle cells induced by platelet-derived growth factor B

Received:2018-07-23 Revised:2018-12-03 Online:2019-02-05 Published:2019-01-16

摘要/Abstract

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PDGF-BB+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a-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a-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。

关键词: 关键词：miR-26a, 血管平滑肌细胞, 表型转换, PDGF-BB

Abstract: Object 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, which were 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 also. Results Compare 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.

Key words: Key word: miR-26a, VSMCs, phenotypic switch, PDGF-BB

洪新宝王炳才余江水杨晓燕郭榕顾信建李月婷. miR-26a在血小板源性生长因子B诱导血管平滑肌细胞表型转换的作用[J]. 基础医学与临床, 2019, 39(2): 192-196.

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