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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 regulatory effect of microRNA-214 (mi R-214) on cardiomyocyte hypertrophy and its possible target genes. Methods: The ventricular myocyte hypertrophy model induced by angiotensin Ⅱ (Ang-Ⅱ) in C57BL / 6 mice was established. The dual luciferase reporter assay was used to detect the expression of mi R-214 and the potential target gene MEF2C 3 ’ (3’UTR). The mRNA and protein levels of MEF2C and hypertrophic markers were detected by real-time quantitative PCR (RT-qPCR) and Western blot respectively. Results: The expressions of cardiac hypertrophy markers ANP, ACTA1 and β-MHC, and mi R-214 were significantly increased in Ang-II-induced hypertrophic mouse cardiomyocytes. Dual luciferase reporter assay suggested that mi R-214 and MEF2C 3’UTR interaction confirmed that mi R-214 can inhibit the expression of MEF2C at the transcriptional level, MEF2C protein levels were significantly upregulated in hypertrophic cardiomyocytes; Overexpression of mi R-214 and silencing MEF2C can inhibit Ang- Ⅱ induced hypertrophy markers in cardiomyocytes. Conclusion: MEF2C is a target gene of mi R-214 and mediates the effect of mi R-214 in inhibiting cardiomyocyte hypertrophy.