本文選題：循環(huán)miRNAs　切入點：非增殖性糖尿病視網(wǎng)膜病變　出處：《寧波大學》2017年碩士論文　論文類型：學位論文

【摘要】：研究背景:糖尿病視網(wǎng)膜病變(Diabetic retinopathy,DR)是一種嚴重的眼科疾病,是糖尿病中主要的并發(fā)癥之一,該疾病主要的特征是視網(wǎng)膜微血管功能障礙,最嚴重的后果是導致失明,是導致糖尿病患者視力喪失的主要因素。MicroRNA(miRNA)是非編碼單鏈微小RNA分子,約為21-24個核苷酸的長度,大量研究表明循環(huán)mi RNAs是一類具有臨床診斷價值的新型生物標志物,目前已有許多循環(huán)mi RNAs用于疾病診斷生物標志物研究的成功案例,而循環(huán)mi RNAs與糖尿病性非增殖性視網(wǎng)膜病變發(fā)生發(fā)展關系尚未明確。目的:獲取在2型糖尿病非增殖性視網(wǎng)膜病變(non-proliferative diabetic retinopathy,NPDR)發(fā)展進程中差異表達的循環(huán)mi RNAs,大樣本驗證篩選出來的循環(huán)mi RNAs,通過(Receiver operating characteristic curve,ROC)曲線預測循環(huán)mi RNAs對2型糖尿病非增殖性視網(wǎng)膜病變的診斷價值,為合理評價循環(huán)mi RNAs作為2型糖尿病非增殖性視網(wǎng)膜病變診斷與治療的生物標志物提供可靠的依據(jù)。方法:本研究所有樣本都是來自深圳市南山區(qū)慢性病防治院,其中健康組血清樣本144例、糖尿病前期(prediabetic)組血清樣本70例、2型糖尿病(type2 diabetes mellitus,T2DM)血清樣本117例及2型糖尿病非增殖性視網(wǎng)膜病變血清樣本75例,2型糖尿病非增殖性視網(wǎng)膜病變經(jīng)眼底熒光血管造影檢查證實。首先運用基因芯片技術初步篩選健康組、糖尿病前期組、2型糖尿病組及2型糖尿病非增殖性視網(wǎng)膜病變組中血清mi RNAs表達譜,每組分別提供樣本5例,根據(jù)芯片結果篩選出表達差異明顯、可能與2型糖尿病非增殖性視網(wǎng)膜病變的進展相關的循環(huán)mi RNAs,并采用實時熒光定量PCR(quantitative real-time PCR,qRTPCR)技術對不同階段差異表達顯著的循環(huán)mi RNAs進一步擴大樣本驗證,以尋找與2型糖尿病非增殖性視網(wǎng)膜病變疾病發(fā)展進程相關的循環(huán)mi RNAs。結果:通過對芯片檢測結果進行篩選,最終發(fā)現(xiàn)在2型糖尿病非增殖性視網(wǎng)膜病變組及2型糖尿病組中有4個miRNAs(miR-3197、miR-2116-5P、miR-3939、miR-1910-3P)表達差異顯著,而在糖尿病前期組、健康組及2型糖尿病組間未發(fā)現(xiàn)有表達差異明顯的mi RNAs。進一步運用q RT-PCR技術擴大樣本量進行驗證,結果顯示miR-3197和miR-2116-5P在2型糖尿病非增殖性視網(wǎng)膜病變組中的表達水平明顯比2型糖尿病組高,P值小于0.001,有統(tǒng)計學意義,且受試者工作特征ROC曲線下面積(Area under curve,AUC)分別為0.967和0.760。結論:循環(huán)miR-2116-5p及循環(huán)miR-3197在2型糖尿病非增殖性視網(wǎng)膜病變患者中表達水平明顯高于2型糖尿病組,與芯片結果一致,提示循環(huán)miR-2116-5p及循環(huán)miR-3197有望作為2型糖尿病非增殖性視網(wǎng)膜病變早期診斷的生物標志物。
[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.
【學位授予單位】：寧波大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R587.2;R774.1

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