文章摘要
张紫微,杨丽霞,郭瑞威,吕晋琳,陆霓虹,王先梅,石燕昆.miR-155抑制AngⅡ诱导的主动脉血管平滑肌细胞周期转换[J].南京医科大学学报,2017,(3):281~286,334
miR-155抑制AngⅡ诱导的主动脉血管平滑肌细胞周期转换
Inhibition of miR-155 on AngⅡ-stimulated cell cycle progression of vascular smooth muscle cells
投稿时间:2016-07-11  
DOI:10.7655/NYDXBNS20170305
中文关键词: miR-155  血管紧张素Ⅱ;血管平滑肌细胞;细胞周期
英文关键词: miR-155  AngⅡ  vascular smooth muscle cell  cell cycle
基金项目:国家自然科学基金面上项目(81170250)
作者单位
张紫微 昆明医科大学昆明总医院临床学院云南 昆明 650500 成都军区昆明总医院心内科云南 昆明 650032 
杨丽霞 成都军区昆明总医院心内科云南 昆明 650032 
郭瑞威 成都军区昆明总医院心内科云南 昆明 650032 
吕晋琳 昆明医科大学昆明总医院临床学院云南 昆明 650500 成都军区昆明总医院心内科云南 昆明 650032 
陆霓虹 昆明医科大学昆明总医院临床学院云南 昆明 650500 成都军区昆明总医院心内科云南 昆明 650032 
王先梅 成都军区昆明总医院心内科云南 昆明 650032 
石燕昆 成都军区昆明总医院心内科云南 昆明 650032 
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中文摘要:
      目的: 观察转染miR-155后对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导小鼠主动脉血管平滑肌细胞(vascular smooth muscle cell,VSMC)细胞周期转换的影响并探讨其机制。方法:原代培养小鼠VSMC,用1×10-6 mol/L AngⅡ作用于VSMC 48 h后,采用实时荧光定量PCR(quantitative real-time PCR,q-RT-PCR)检测空白对照组及AngⅡ组miR-155表达水平。用q-RT-PCR及Western blot检测分别转染miR-155及阴性对照后各组AngⅡ 1型受体(angiotensin Ⅱ type 1 receptor,AT1R)的mRNA及蛋白表达水平;用Western blot检测转染miR-155 mimic及阴性对照后对AT1R下游细胞外信号调节激酶1/2(extracellular signal-regulated kinase 1/2,ERK1/2)、核糖体蛋白S6激酶(P70S6K1)通路的影响。分别检测转染miR-155 mimic及使用血管紧张素受体阻滞剂缬沙坦、mTOR通路抑制剂雷帕霉素、ERK1/2抑制剂U0126对细胞周期素D1(Cyclin D1)的表达影响,并使用流式细胞分析检测细胞周期变化,探讨miR-155对细胞周期的影响及其机制。结果:AngⅡ可显著降低miR-155的表达。miR-155可从mRNA及蛋白水平抑制AT1R表达,并抑制AngⅡ促AT1R表达的作用。转染miR-155 mimic后可显著抑制AngⅡ促进ERK1/2、P70S6K1通路激活的作用。转染miR-155 mimic、使用缬沙坦、雷帕霉素、U0126抑制AngⅡ促Cyclin D1表达的作用,并使细胞周期阻滞在G0/G1期。结论:miR-155可通过抑制AT1R间接抑制AngⅡ促进ERK1/2、P70S6K1通路激活及Cyclin D1表达的作用,抑制AngⅡ促进细胞周期转换的作用。
英文摘要:
      Objective: To investigate the effect of miR-155 on cell cycle progression of mice vascular smooth muscle cells under treatment of AngⅡ, and explore the detailed mechanism. Methods: The vascular smooth muscle cells(VSMCs) derived from C57 mice were cultured by the adherent method (1×10-6 mol/L AngⅡ for 48 h). Quantitative real-time PCR (q-RT-PCR) was performed to detect the miR-155 expression levels of the blank control group and the Ang Ⅱ group. q-RT-PCR and Western blot assay were performed to detect the mRNA and protein levels of angiotensin Ⅱ type 1 receptor(AT1R) in the transfected miR-155 group and the negative control group; Western blot assay was performed to detect the effect of transfected miR-155 mimics and negative control group on extracellular signal-regulated kinase 1/2 (ERK1/2) of AT1R downstream and ribosomal protein S6 kinase (P70S6K1) signaling pathway. The expression of cyclin D1(Cyclin D1) was examined by transfection of miR-155 mimics,angiotensin receptor blocker valsartan,mTOR pathway inhibitor rapamycin,and ERK1/2 inhibitor U0126. In the end,we used flow cytometer to analyze the change of cell cycle to investigate the effect of miR-155 on cell cycle and its mechanism. Results: AngⅡ significantly decreased the expression of miR-155. miR-155 mimics remarkably attenuated AT1R expression from mRNA and protein levels,inhibited the AngⅡ-activated ERK1/2 and P70S6K signaling pathway,and decreased the AngⅡ-enforced expression of Cyclin D1. Transfection of miR-155,valsartan,rapamycin and U0126 suppressed the AngⅡ-induced G1-to S-phase progression of VSMC. Conclusion: MiR-155 can inhibit the expression of AngⅡ-induced ERK1/2,activation of P70S6K1 signaling pathway,and Cyclin D1 by blocking AT1R,and thereby inhibit the function of Ang Ⅱ-induced cell cycle transition.
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