摘要
目的:研究微小RNA-214(mi R-214)对心肌细胞肥大的调控作用及其可能的作用靶基因。方法:建立血管紧张素Ⅱ(angiotensin-Ⅱ,Ang-Ⅱ)诱导的C57BL/6乳小鼠心室肌细胞肥大模型;双萤光素酶报告基因实验检测mi R-214与潜在靶基因MEF2C 3’端非翻译区(3’UTR)的结合作用;实时荧光定量PCR(RT-q PCR)和Western blot法分别检测MEF2C及肥厚标志物的m RNA和蛋白表达水平。结果:心肌肥厚标志物ANP、ACTA1和β-MHC,以及mi R-214的表达在Ang-II诱导肥大的小鼠心肌细胞中显著增强;双萤光素酶报告基因实验提示mi R-214与MEF2C 3’UTR相互作用,证实mi R-214可在转录水平抑制MEF2C的表达,MEF2C蛋白水平在肥大的心肌细胞中显著上调;过表达mi R-214及沉默MEF2C均能一致性地抑制Ang-Ⅱ诱导的心肌细胞中肥大标志物的表达。结论:MEF2C是mi R-214的靶基因,并介导了mi R-214发挥抑制心肌细胞肥大的作用。
AIM： To investigate the effect of micro RNA-214（miR-214） on cardiomyocyte hypertrophy and the expression of the potential target genes. METHODS： A cell model of hypertrophy was established based on angiotensin-Ⅱ（Ang-Ⅱ）-induced neonatal mouse ventricular cardiomyocytes（NMVCs）. Dual luciferase reporter assay was performed to verify the interaction between mi R-214 and the 3 ＇UTR of MEF2 C. The expression of MEF2C and hypertrophy-related genes at m RNA and protein levels was determined by RT-q PCR and Western blot,respectively. RESULTS： The expression of ANP,ACTA1,β-MHC and mi R-214 was markedly increased in Ang-Ⅱ-induced hypertrophic cardiomyocytes. Dual luciferase reporter assay revealed that mi R-214 interacted with the 3 ＇UTR of MEF2C,and mi R-214 was verified to inhibit MEF2C expression at the transcriptional level. The protein expression of MEF2C was markedly increased in the hypertrophic cardiomyocytes. Moreover,mi R-214 mimic,in parallel to MEF2C si RNA,inhibited the expression of hypertrophy-related genes in Ang-Ⅱ-induced NMVCs. CONCLUSION： MEF2 C is a target gene of mi R-214,which mediates the effect of miR-214 on attenuating cardiomyocyte hypertrophy.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2016年第8期1345-1350,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81270222
No.81470439)
广东省自然科学基金资助项目(No.2014A030313635)
广东省医学研究基金资助项目(No.A2015187)
