【摘要】 目的 筛选并验证T2DM患者相关血浆外泌体微小RNA(mi RNA)，评价差异mi RNA对T2DM的预测能力。方法 选取“河南农村队列”2020年驻马店现场随访的T2DM患者52例及同期非DM者52例，采用三阶段1:1匹配为发现集T2DM(DT2DM,n=5)组、发现集对照（DCon ,n=5）组、验证集T2DM(VT2DM,n=15)组、验证集对照（VCon,n=15）组、靶向验证集T2DM(TT2DM,n=32)组和靶向验证集对照（TCon,n=32）组。采用外泌体mi RNA测序技术对血浆外泌体mi RNA进行测序，q RT-PCR检测关键差异mi RNA水平，Spearman相关分析mi RNA与FPG、FIns的相关性，Logistic回归分析mi RNA对T2DM的影响，受试者工作特征（ROC）曲线评价关键mi RNA对T2DM的预测价值。结果 DT2DM组心率、FPG、外泌体浓度高于DCon组（P<0.05）。VT2DM组TG、FPG、FIns、腹型肥胖比例高于VCon组（P<0.05）。TT2DM组WC、TG、FPG、高血压病比例高于TCon组（P<0.05）。两阶段测序结果显示，DT2DM、VT2DM组外泌体关键mi RNA miR-3120-5p表达水平均高于DCon、VCon组。Spearman相关分析显示，校正混杂因素后，mi R-3120-5p与FPG呈正相关（r=0.311,P=0.016）。Logistic回归分析显示，校正混杂因素后，外泌体mi R-3120-5p表达水平是T2DM的影响因素（OR 1.566,95%CI 1.177～2.057）。ROC曲线分析显示，外泌体mi R-3120-5p诊断T2DM的曲线下面积为0.75，敏感度为81%，特异度为66%，截断值为7.28。结论 miR-3120-5p在T2DM患者血浆外泌体中高表达，并对其具有良好的预测价值，但需在大样本多中心队列人群中进一步验证。更多还原

【Abstract】 Objective To identify and validate the plasma-derived exosomal microRNA(mi RNA) associated with type 2 diabetes mellitus(T2DM),and to assess the predictive ability of differential miRNA for T2DM. Methods 52 newly diagnosed T2DM patients enrolled from the “Henan rural cohort” in Zhumadian in 2020 and 52 non-diabetic individuals under going physical examinations during the same period were included. Using a three-stage 1:1 matching design, the following groups were formed:discovery set T2DM(DT2DM,n=5)group,discovery set control(DCon,n=5)group,validation set T2DM(VT2DM,n=15)group, validation set control(VCon,n=15)group,targeted validation set T2DM(TT2DM,n=32)group,and targeted validation set control(TCon,n=32)group. Plasma-derived exosomal miRNA was sequenced using the exosomal miRNA sequencing technology. Key differentially expressed miRNAs were quantified by q RT-PCR. Spearman correlation analysis was performed to examine the association between miRNAs and fasting plasma glucose(FPG) and fasting insulin(FIns). Logistic regression was used to evaluate the influence of miRNAs on T2DM. The predictive performance of key miRNAs for T2DM was evaluated by the receiver operating characteristic(ROC) curve. Results Heart rate, FPG, and exosome concentration were higher in the DT2DM group than in the DCon group(P<0. 05). Triglyceride(TG), FPG, FIns, and proportion of abdominal obesity were higher in the VT2DM group than in the VCon group(P<0. 05). Waist circumference,TG,FPG,and proportion of hypertension were higher in the TT2DM group than in the TCon group(P<0. 05). Two-stage sequencing revealed that expression of the key miRNA miR-3120-5p in exosomes were higher in the DT2DM and VT2DM groups than in the DCon and VCon groups. After adjusting for confounding factors,Spearman correlation analysis showed a positive correlation between mi R-3120-5p and FPG(r=0. 311,P=0. 016). Logistic regression analysis revealed that the expression level of exosomal miR-3120-5p was an influencing factor for T2DM after adjusting for confounding factors(OR 1. 566,95% CI 1. 177～2. 057). The ROC curve analysis showed that the area under the curve for exosomal miR-3120-5p in diagnosing T2DM was 0. 75,with a sensitivity of 81%,a specificity of 66%,and a cut-off value of 7. 28. Conclusions The miR-3120-5p is highly expressed in plasma exosomes and has good predictive value for T2DM. Further validation in multicenter cohorts is warranted.更多还原