本文選題：2型糖尿病 + 微血管并發(fā)癥　； 參考：《南京師范大學(xué)》2015年碩士論文

【摘要】：目的：篩選2型糖尿病(Type 2 diabetes mellitus, T2DM)及其微血管并發(fā)癥(Type 2 diabetes mellitus microvascular complications,T2DMC)患者特異變化血清微小核糖核酸(microRNA,miRNA)表達(dá)譜,評(píng)價(jià)其對(duì)T2DM及T2DMC的診斷、鑒別診斷和病情監(jiān)控臨床價(jià)值。方法：收集2013年9月～2014年5月間南京軍區(qū)總院內(nèi)分泌科收治的新發(fā)或一個(gè)月內(nèi)未經(jīng)任何治療的T2DM及T2DMC各87例,同時(shí)收集同期在醫(yī)院體檢中心年齡、性別完全匹配的健康體檢者87例做為正常對(duì)照。首先采用高通量、高靈敏度的低密度芯片技術(shù)(TaqMan Low Density Array, TLDA)對(duì)T2DM.T2DMC及健康對(duì)照各25例樣本分別組成的混合血清中768種miRNA的表達(dá)水平進(jìn)行檢測(cè),初篩T2DM和T2DMC特異變化的一組血清miRNA;然后運(yùn)用準(zhǔn)確的實(shí)時(shí)熒光定量PCR技術(shù)(quantitative real-time polymerase chain reaction, qRT-PCR)對(duì)初篩miRNA在TLDA所使用的同一批樣本中進(jìn)行復(fù)篩(復(fù)篩組)；進(jìn)一步運(yùn)用qRT-PCR技術(shù)在另外三組T2DM.T2DMC和健康對(duì)照各62例樣本中對(duì)復(fù)篩結(jié)果進(jìn)行驗(yàn)證(驗(yàn)證組),篩選T2DM及T2DMC患者血清中穩(wěn)定變化的一組miRNA。同時(shí),運(yùn)用酶聯(lián)免疫吸附法(enzyme-linked immunosorbent assay, ELISA)測(cè)定全部患者及對(duì)照血清樣本中氧化低密度脂蛋白(ox-LDL).氧化脂蛋白(a) [ox-Lp(a)]、β2糖蛋白氧化低密度脂蛋白復(fù)合物(p2-GPI-ox-LDL)和β2糖蛋白脂蛋白(a)復(fù)合物[β2-GPI-Lp(a)]勺水平,運(yùn)用Spearman秩相關(guān)分析和邏輯回歸分析患者特異變化血清miRNA與上述脂蛋白、臨床指標(biāo)相關(guān)性及是否為T2DM及T2DMC獨(dú)立危險(xiǎn)因素。結(jié)果：TLDA結(jié)果顯示,T2DM及T2DMC患者血清miRNA表達(dá)譜與健康對(duì)照相比具有明顯差異,53種miRNA在T2DM及T2DMC患者血清中較對(duì)照組表達(dá)上升,117種miRNA在患者表達(dá)下降。結(jié)合TLDA初篩及文獻(xiàn)調(diào)查結(jié)果,我們選取了12種miRNA在復(fù)篩組樣本中進(jìn)行復(fù)篩,發(fā)現(xiàn)8種miRNA,包括miR-571、miR-661、 miR-1303、miR-152、miR-1323、miR-892b、miR-770-5p和miR-886-3p水平在兩組患者血清中顯著上升,其中在T2DMC患者中變化更為顯著(變化倍數(shù)2,P0.05)；8種miRNA在驗(yàn)證組中變化趨勢(shì)與復(fù)篩組一致(變化倍數(shù)2,P0.05)；ROC曲線分析顯示,8種血清miRNA對(duì)T2DM診斷的ROC曲線下面積(AUC)在0.774～0.877之間,對(duì)T2DMC診斷的ROC曲線下面積在0.823～0.946之間,對(duì)T2DM和T2DMC鑒別診斷的ROC曲線下面積在0.611～0.675之間。根據(jù)選定的cutoff值,8種血清miRNA對(duì)T2DM診斷的靈敏性和特異性分別在68.2%～88.1%和63.0%～84.5%之間；對(duì)T2DMC診斷的靈敏性和特異性分別在64.8%～84.7%和80.6%～98.6%之間；進(jìn)一步分析上述miRNA的鑒別診斷價(jià)值顯示,3種miRNA對(duì)T2DM和T2DMC的鑒別診斷具一定臨床價(jià)值,其AUC、靈敏性和特異性分別為：miR-886-3p(AUC=0.675,靈敏性=0.651,特異性=0.628)、miR-892b(AUC=0.639,靈敏性=0.662,特異性=0.622)和miR-571(AUC=0.611,靈敏性=0.603,特異性=0.608)。ELISA結(jié)果顯示,T2DM和T2DMC患者ox-LDL、ox-Lp(a)、β2-GPI-ox-LDL、β2-GPI-Lp(a)水平亦顯著高于健康對(duì)照組(P均0.05)。Spearman秩相關(guān)分析顯示,T2DM和T2DMC患者變化的8種血清miRNA與空腹血糖(fasting blood-glucose, FPG)、ox-LDL、ox-Lp(a)呈顯著正相關(guān)(P均0.05)。單因素邏輯回歸分析顯示,8種血清miRNA、氧化脂蛋白及其復(fù)合物均為預(yù)測(cè)T2DM和T2DMC患者發(fā)病的獨(dú)立危險(xiǎn)因素；進(jìn)一步多因素分析結(jié)果顯示,在排除性別、年齡等影響因素后,miR-770、miR-661和甘油三酯仍是兩組患者獨(dú)立危險(xiǎn)因素。結(jié)論：8種血清包括miR-571、miR-661、miR-1303、miR-152、miR-1323、miR-892b、 miR-770-5p和miR-886-3p在T2DM和T2DMC患者患者中顯著上調(diào),有望作為T2DM及T2DMC的輔助診斷標(biāo)志物。
[Abstract]:Objective : To investigate the expression profiles of microribonucleic acid ( microRNA , miRNA ) in patients with type 2 diabetes mellitus ( T2DM ) and its microvascular complications ( Type 2 diabetes mellitus complicated complications , T2DMC ) .
Serum levels of low density lipoprotein ( ox - LDL ) in sera of patients with T2DM and T2DMC were determined by using the enzyme - linked immunosorbent assay ( ELISA ) . Results : The results showed that the levels of serum miRNA in patients with T2DM and T2DMC were significantly higher than those in healthy controls . Results : The results showed that the levels of serum miRNA in patients with T2DM and T2DMC were significantly higher than those in healthy controls .
The change trend of 8 kinds of miRNA in the validation group was consistent with that of the compound sieve group ( changing multiple 2 , P0.05 ) .
ROC curve analysis showed that the area under ROC curve ( AUC ) of 8 kinds of serum miRNA in the diagnosis of T2DM was between 0.784 and 0.877 , the area under ROC curve of T2DMC diagnosis was between 0.823 and 0.694 , and the area under ROC curve for the differential diagnosis of T2DM and T2DMC was 0.611 - 0.675 . According to the selected cutoff value , the sensitivity and specificity of 8 kinds of serum miRNA to the diagnosis of T2DM were 68.2 % - 88.1 % and 63.0 % - 84.5 % , respectively ;
The sensitivity and specificity of T2DMC diagnosis were 64.8 % 锝，

本文編號(hào)：2061330