代谢综合征人群尿液miRNA-29a-3p的表达变化及其与代谢亚组分的相关性研究

发布时间：2018-05-20 16:48

本文选题：循环microRNA + miR-29a-3p　；参考：《浙江大学》2015年硕士论文

【摘要】：[目的和背景] 代谢综合征(metabolic syndrome,MetS)是中心性肥胖、糖代谢异常(糖尿病或糖调节受损)、高血压和血脂异常等多种疾病或危险因素在个体聚集的一组临床征候群。目前认为其病理生理基础可能是胰岛素抵抗,但具体机制仍不确切。近年来,关于microRNA(简称miRNA)的发现和研究异军突起,miRNA被证实参与和调节人类生理病理的多个环节。现已有多项证据表明,循环miRNAs可能参与和影响糖脂代谢。本课题通过测定miRNA-29a-3p(以下简称miR-29a-3p)在代谢综合征人群尿液中的表达量,观察miR-29a-3p与各代谢组分的相关性,评估其患病风险和诊断价值,希望能给代谢综合征的诊断和治疗带来全新的思路和方法。 [方法] 将代谢综合征患者及正常对照组共8例血清样本进行了基于377个miRNAs探针的TaqMan MicroRNA芯片检测,发现11个异常表达的miRNAs,其中7个为高表达,4个为低表达。本实验选取其中高表达的miR-29a-3p作为研究对象,运用qRT-PCR技术在40例MetS及40例非MetS人群的尿液样本中进行了验证,通过t检验比较了两组尿液miR-29a-3p的表达差异,并进一步运用Spearson相关,多元逐步线性回归,多元logistics回归分析,ROC曲线等统计学方法对miR-29a-3p与各代谢亚组分进行相关性、患病风险和诊断价值的分析。 [结果] TaqMan MicroRNA芯片检测结果显示代谢综合征人群存在11个异常表达的miRNAs,其中有7个为高表达,分别是miR-20a-3p、miR-143a-3p、miR-365a-3p、 miR-22a-5p、miR-71a-5p、miR-29a-3p、miR-99a-5p、进一步对其中miR-29a-3p行qRT-PCR验证,MetS组尿液miR-29a-3p的表达水平比非MetS组显著升高(p0.001)；多元逐步线性回归结果显示尿液miR-29a-3p与空腹胰岛素(beta=0.561, p0.001)、高密度脂蛋白(beta=0.242,p0.001)和BMI(beta=-0.141,p=0.025)成独立相关。将尿液miR-29a-3p的qRT-PCR检测所得Ct值计算相对浓度后进行二分组(分组界值为0.96),并控制年龄、性别、吸烟和饮酒因素,结果发现相对浓度大于0.96组,其MetS的患病风险率是浓度小于0.96组的4.01倍(95%CI:1.22-13.12; p=0.022)。ROC曲线结果提示：尿液miR-29a-3p的表达水平对MetS具有一定诊断价值(AUC:0.776,P=0.001;95%CI:0.603-0.829),其诊断敏感性77.5%,特异性60%。 [结论] 尿液miR-29a-3p在MetS人群中相比非MetS人群其表达显著增高,它可能通过促糖脂代谢异常来介导MetS的发生。尿液miR-29a-3p表达水平对MetS具有一定的诊断价值,有望成为新的分子诊断标志物。这为进一步研究miR-29a-3p介导MetS发生的机制和生物学行为提供理论依据,为MetS的靶向治疗提供全新思路。
[Abstract]:[purpose and background] Metabolic syndrome (MS) is a group of clinical symptoms of central obesity, abnormal glucose metabolism (diabetes mellitus or impaired glucose regulation, hypertension and dyslipidemia). It is believed that insulin resistance may be the pathophysiological basis of insulin resistance, but the mechanism is still unclear. In recent years, the discovery and research of microRNAs (miRNAs) have been proved to be involved in and regulate human physiology and pathology. There is evidence that circulating miRNAs may be involved in and affect glucose and lipid metabolism. In this study, we measured the expression of miRNA-29a-3p (miR-29a-3p) in the urine of patients with metabolic syndrome, observed the correlation between miRNA-29a-3pand various metabolites, and evaluated the risk and diagnostic value of miRNA-29a-3p. We hope to bring new ideas and methods to the diagnosis and treatment of metabolic syndrome. [methods] A total of 8 serum samples from patients with metabolic syndrome and normal control were detected by TaqMan MicroRNA microarray based on 377 miRNAs probes. 11 abnormal expression of miRNas were found, including 7 high expression and 4 low expression. In this experiment, the high expression of miR-29a-3p was selected as the research object, and the urine samples of 40 cases of MetS and 40 cases of non-MetS were tested by qRT-PCR technique. The difference of the expression of miR-29a-3p in urine between the two groups was compared by t test, and Spearson correlation was used further. Multivariate stepwise linear regression and multivariate logistics regression analysis were used to analyze the correlation between miR-29a-3p and various metabolic subcomponents, the risk of disease and the diagnostic value. [results] The results of TaqMan MicroRNA microarray analysis showed that there were 11 abnormal expression of miRNas in metabolic syndrome population, 7 of which were overexpression. They were miR-20a-143a-3pnmiR-365a-3p, miR-22a-5pnmiR-71a-5pnmiR-29a-3pnmiR-99a-5p. the level of urinary miR-29a-3p in miR-29a-3p group was significantly higher than that in non-MetS group, and the multivariate stepwise linear regression analysis showed that urine miR-29a-3p was correlated with fasting insulin beta1 0.561, p0.001, high density lipoprotein beta1 0.242p0.001) and BMIbetaI 0.141p0. 025) respectively. The results of multiple stepwise linear regression showed that there was an independent correlation between urine miR-29a-3p and fasting insulin Beta 0.561, p0.001, high density lipoprotein beta1 0.242p0.001) and BMIbeta- 0.141 p0. 025). After calculating the relative concentration of Ct by qRT-PCR detection of urine miR-29a-3p, the group was divided into two groups (the threshold of the group was 0.96) and the factors of age, sex, smoking and drinking were controlled. The results showed that the relative concentration was greater than 0.96. The risk rate of MetS was 4.01 times of 95 CI: 1.22-13.12 in 0.96 group. The results of p=0.022).ROC curve indicated that the level of urinary miR-29a-3p expression had certain diagnostic value for MetS. The diagnostic sensitivity was 77.5% and specificity was 605%. [conclusion] The expression of miR-29a-3p in urine was significantly higher in MetS than in non-MetS, and it may mediate the occurrence of MetS by abnormal glucose and lipid metabolism. Urine miR-29a-3p expression level has certain diagnostic value to MetS, and it is expected to be a new molecular diagnostic marker. This provides a theoretical basis for the further study of the mechanism and biological behavior of MetS mediated by miR-29a-3p, and provides a new idea for the targeted treatment of MetS.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R589

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