摘要
目的:通过检测miR181b在心肌肥厚患者外周血中的表达,研究miR181b与PRKG-1在心肌肥大中的作用机制,为心肌肥厚临床诊断和治疗提供新的靶点。方法:采用实时定量PCR方法检测50例临床诊断为心肌肥厚患者及25例正常健康人的外周血miR181b的水平,分析其表达水平与临床病理特征的关系。结果:miR181b在心肌肥厚患者外周血中表达(3.35±0.22)升高,差异有统计学意义(P<0.05);体外大鼠原代心肌细胞在镜下呈多角形,采用α-横纹肌肌动蛋白抗体和DAPI进行免疫荧光染色,镜下90%以上细胞发出绿色荧光,表明原代培养心肌细胞纯度较高;采用去氧肾上腺素处理心肌细胞72 h后,心肌细胞总蛋白表达显著升高(P<0.05);流式细胞术结果显示PE组心肌细胞大小明显增加(P<0.05);实时定量PCR检测结果表明,PE组中心肌肥大相关基因β-MHC、α-SA、ANP显著升高(P<0.05),证明体外构建心肌肥大模型成功。结论:miR181b在心肌肥厚患者外周血中表达显著上调;体外成功培养乳鼠原代心肌细胞,通过PE处理法成功构建体外心肌肥大模型;在肥大心肌细胞中,miR181b与prkg-1 m RNA及蛋白的表达有显著相关性;miR181b可能通过调控prkg-1 m RNA及蛋白水平的表达参与心肌肥大,有望成为心肌肥厚临床诊断和治疗的新的靶点。
Objective： By detecting miR181b in expression of periphera｜ blood in the patients with myocardial hypertrophy, mechanism research and miR181b PRKG-1 in myocardial hypertrophy, for clinical diagnosis and treatment of cardiac hypertrophy provide new targets. Method： Using quantitative real-time PCR assay for the detection of 50 cases of clinical diagnosis for the level of peripheral blood miR181b patients with hypertrophic cardiomyopathy （ HCM ） and 25 normal healthy patients, to analyze the relationship between the expression level and clinicopathological characteristics. Result： The expression of miR181b in peripheral blood of patients with myocardial hypertrophy （ 3.35 ± 0.22 ） increased, the difference was statistically significant （ P〈0.05 ）; in vitro primary rat myocardial cells were polygonal in microscope, using alpha sarcomeric actin antibody and DAPI immunofluorescence staining, microscope 90% above cells emitted green fluorescence the results showed that the cell, high purity of primary cultured myocardial ceils; using the phenylephrine treatment after 72 h, the expression of the total protein of myocardial cells significantly increased （ P〈0.05 ） ; flow cytometry showed that PE group myocardial cell size increased significantly （ P〈0.05 ） ; real time quantitative PCR test results showed that, PE group of Central muscle hypertrophy related genes of beta -MHC -SA, ANP, alpha significantly increased （P〈0.05）, that in vitro cardiac hypertrophy model. Conclusion： miR181b in peripheral blood of patients with myocardial hypertrophy expression is significantly up-regulated; primary cultured myocardial cells of neonatai rat culture successfully in vitro, through PE treat successfully construct in vitro model of cardiac hypertrophy; in cardiac myocyte hypertrophy, miR181b and prkg-1 mRNA and protein expression have significant correlation; miR181b may by regulation prkg-1 mRNA and protein level expression in cardiac hypertrophy, is expected to become a new target for diagnosis and treatment of myocardial hypertrophy.
出处
《中国医学创新》
CAS
2016年第1期8-11,共4页
Medical Innovation of China
