循环miRNAs与2型糖尿病非增殖性视网膜病变的相关性研究

发布时间：2018-03-03 08:37

本文选题：循环miRNAs　切入点：非增殖性糖尿病视网膜病变　出处：《宁波大学》2017年硕士论文　论文类型：学位论文

【摘要】：研究背景:糖尿病视网膜病变(Diabetic retinopathy,DR)是一种严重的眼科疾病,是糖尿病中主要的并发症之一,该疾病主要的特征是视网膜微血管功能障碍,最严重的后果是导致失明,是导致糖尿病患者视力丧失的主要因素。MicroRNA(miRNA)是非编码单链微小RNA分子,约为21-24个核苷酸的长度,大量研究表明循环mi RNAs是一类具有临床诊断价值的新型生物标志物,目前已有许多循环mi RNAs用于疾病诊断生物标志物研究的成功案例,而循环mi RNAs与糖尿病性非增殖性视网膜病变发生发展关系尚未明确。目的:获取在2型糖尿病非增殖性视网膜病变(non-proliferative diabetic retinopathy,NPDR)发展进程中差异表达的循环mi RNAs,大样本验证筛选出来的循环mi RNAs,通过(Receiver operating characteristic curve,ROC)曲线预测循环mi RNAs对2型糖尿病非增殖性视网膜病变的诊断价值,为合理评价循环mi RNAs作为2型糖尿病非增殖性视网膜病变诊断与治疗的生物标志物提供可靠的依据。方法:本研究所有样本都是来自深圳市南山区慢性病防治院,其中健康组血清样本144例、糖尿病前期(prediabetic)组血清样本70例、2型糖尿病(type2 diabetes mellitus,T2DM)血清样本117例及2型糖尿病非增殖性视网膜病变血清样本75例,2型糖尿病非增殖性视网膜病变经眼底荧光血管造影检查证实。首先运用基因芯片技术初步筛选健康组、糖尿病前期组、2型糖尿病组及2型糖尿病非增殖性视网膜病变组中血清mi RNAs表达谱,每组分别提供样本5例,根据芯片结果筛选出表达差异明显、可能与2型糖尿病非增殖性视网膜病变的进展相关的循环mi RNAs,并采用实时荧光定量PCR(quantitative real-time PCR,qRTPCR)技术对不同阶段差异表达显著的循环mi RNAs进一步扩大样本验证,以寻找与2型糖尿病非增殖性视网膜病变疾病发展进程相关的循环mi RNAs。结果:通过对芯片检测结果进行筛选,最终发现在2型糖尿病非增殖性视网膜病变组及2型糖尿病组中有4个miRNAs(miR-3197、miR-2116-5P、miR-3939、miR-1910-3P)表达差异显著,而在糖尿病前期组、健康组及2型糖尿病组间未发现有表达差异明显的mi RNAs。进一步运用q RT-PCR技术扩大样本量进行验证,结果显示miR-3197和miR-2116-5P在2型糖尿病非增殖性视网膜病变组中的表达水平明显比2型糖尿病组高,P值小于0.001,有统计学意义,且受试者工作特征ROC曲线下面积(Area under curve,AUC)分别为0.967和0.760。结论:循环miR-2116-5p及循环miR-3197在2型糖尿病非增殖性视网膜病变患者中表达水平明显高于2型糖尿病组,与芯片结果一致,提示循环miR-2116-5p及循环miR-3197有望作为2型糖尿病非增殖性视网膜病变早期诊断的生物标志物。
[Abstract]:Background: diabetic retinopathy (DRR) is a severe ophthalmic disease and one of the major complications of diabetes mellitus. It is characterized by retinal microvascular dysfunction, the most serious of which is blindness. MicroRNAs miRNAs are the main factors leading to vision loss in diabetic patients. MicroRNAs are non-coding single-stranded microRNAs, which are about 21-24 nucleotides in length. A large number of studies have shown that circulating mi RNAs is a new biomarker with clinical diagnostic value. There have been many successful cases of circulating mi RNAs used in the biomarkers of disease diagnosis. But the relationship between circulatory mi RNAs and diabetic nonproliferative retinopathy is not clear. Objective: to obtain the differential expression of circulatory mi diabetic retinopathy in the development of type 2 diabetic non-proliferative diabetic retinopathy. The value of circulating mi RNAs in the diagnosis of type 2 diabetic non-proliferative retinopathy was predicted by using the receiver operating characteristic curve. To provide a reliable basis for the reasonable evaluation of circulating mi RNAs as a biomarker for the diagnosis and treatment of type 2 diabetic non-proliferative retinopathy. The serum samples of the healthy group were 144 cases. Serum samples of 70 patients with type 2 diabetes mellitus type 2 diabetes mellitus T2DM and 75 patients with type 2 diabetic non-proliferative retinopathy via fundus fluorescein angioplasty. Shadow examination confirmed. First of all, the gene chip technology was used to screen the healthy group. The expression profiles of serum mi RNAs in prediabetic patients with type 2 diabetes mellitus and type 2 diabetic non-proliferative retinopathy were collected from 5 patients in each group, and the difference was significant according to the results of microarray screening. Circulatory mi RNAss, which may be associated with the progression of type 2 diabetic non-proliferative retinopathy, were further expanded to verify the difference in expression of circulatory mi RNAs at different stages by real-time fluorescent quantitative PCR(quantitative real-time rrr qRT PCR. To search for circulatory mi RNAs associated with the progression of type 2 diabetic non-proliferative retinopathy. Results: the results of microarray detection were screened. It was finally found that there were significant differences in the expression of miRNAsP miR-3197 miR-2116-5PmmiR-3939 miR-1910-3Pin type 2 diabetic non-proliferative retinopathy group and type 2 diabetes group. There was no significant difference in the expression of mi RNAs between the healthy group and the type 2 diabetic group. The Q RT-PCR technique was used to expand the sample size. The results showed that the expression of miR-3197 and miR-2116-5P in type 2 diabetic non-proliferative retinopathy group was significantly higher than that in type 2 diabetes mellitus group (P < 0.001). The area area under (AUC) under the operating characteristic ROC curve was 0.967 and 0.760 respectively. Conclusion: the expression of circulating miR-2116-5p and circulating miR-3197 in patients with type 2 diabetic non-proliferative retinopathy is significantly higher than that in type 2 diabetic patients, which is consistent with the results of microarray. These results suggest that circulating miR-2116-5p and circulating miR-3197 may be used as biomarkers for early diagnosis of type 2 diabetic nonproliferative retinopathy.
【学位授予单位】：宁波大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.2;R774.1

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