2型糖尿病及其微血管并發癥患者血清miR-661水平變化及其輔助診斷價值*

王靜，王成，萬淑君，李丹丹，張春妮，汪俊軍

(南京軍區南京總醫院臨床檢驗科，南京 210002)

·臨床實驗研究·

2型糖尿病及其微血管并發癥患者血清miR-661水平變化及其輔助診斷價值*

王靜，王成，萬淑君，李丹丹，張春妮，汪俊軍

(南京軍區南京總醫院臨床檢驗科，南京 210002)

目的觀察2型糖尿病(T2DM)及其微血管并發癥(T2DMC)患者血清miR-661水平變化并分析其臨床輔助診斷價值。方法用實時熒光定量PCR技術(qRT-PCR)檢測60例T2DM患者、60例T2DMC患者及60例健康對照者血清中miR-661水平，同時測定空腹血糖(FPG)、總膽固醇(TC)、三酰甘油(TG)、低密度脂蛋白膽固醇(LDL-C)、高密度脂蛋白膽固醇(HDL-C)和糖化血紅蛋白(HbA1c)等指標水平，ROC曲線、相關性分析及Logistic regression分析血清miR-661對T2DM及T2DMC的診斷價值。結果qRT-PCR結果顯示，T2DM患者和T2DMC患者血清miR-661相對含量[中位數(四分位數)][M(P25，P75)]分別為230.1(83.6，426.2)fmol/L和441.3(212.9，1 021.0)fmol/L，與健康人對照組的118(63.6，192.4)fmol/L相比，均升高(U值分別為1 207和435，P均<0.01)；血清miR-661 T2DM的ROC曲線下面積(AUCROC)為0.665(95%CI=0.565～0.765，P<0.01)，T2DMC的AUCROC為0.879(95%CI=0.821～0.938，P<0.01)，區分T2DM及T2DMC的AUCROC為0.694(95%CI=0.600～0.788，P<0.01)；相關性分析結果顯示血清miR-661水平與FPG和LDL-C呈正相關(r值分別為0.291和0.149，P均<0.05)，與HDL-C呈負相關(r為-0.243，P<0.01)；進一步用單因素和多因素Logistic regression分析結果顯示，血清miR-661是T2DM和T2DMC的獨立危險因素(P均<0.01)。結論血清中表達升高的miR-661是T2DM及T2DMC患者潛在輔助診斷標志物及獨立危險因素。

2型糖尿病；微血管并發癥；微小核糖核酸；血清；生物標志物

Abstract:ObjectiveTo investigate the serum levels of miR-661 in type 2 diabetes(T2DM)patients and type 2 diabetes with microvascular complications(T2DMC), and to further evaluate its clinical auxiliary diagnosis significance.MethodsThe serum levels of miR-661 were examined in 60 T2DM patients, 60 T2DMC patients and 60 healthy controls using quantitative real-time PCR(qRT-PCR). The sera levels of biochemical parameters including fasting plasma glucose(FPG), cholesterol(TC), triglycerides(TG), low density lipoprotein(LDL-C), high density lipoprotein(HDL-C)and haemoglobinA1C(HbA1c)were determined by biochemical analyzer. The ROC curve analysis, correlation analyses and logistic regression analyses were performed to evaluate the clinical auxiliary diagnosis value of serum miR-661, respectively.ResultsThe qRT-PCR results showed that the concentrations of miR-661 was 118(63.6,192.4)fmol/L in healthy controls, which were significant lower than those of T2DM patients[230.1(83.6,426.2)fmol/L,(U=1 207，P<0.01)]and T2DMC patients[441.3(212.9,1 021)fmol/L,(U=435，P<0.01)]. ROC curve(AUCROC)of miR-661 was 0.665( 95%CI: 0.585～0.765,P<0.01)for T2DM patients and 0.879( 95%CI: 0.821 ～ 0.938,P<0.01)for T2DMC patients. The AUCROCbetween T2DM and T2DMC patients was 0.694(95%CI: 0.600～0.788,P<0.01). The correlation analyses revealed that the levels of miR-661 was positively correlated with concentrations of FPG(r=0.291,P<0.05)and LDL-C(r=0.149,P<0.05), and negatively correlated with concentrations of HDL-C(r=-0.243，P<0.01). Logistic regression analysis revealed that serum miR-661 was an independent risk factor in T2DM and T2DMC(P<0.01).ConclusionSerum miR-661 was potential biomarker and risk factor for T2DM and T2DMC.

Keywords: type 2 diabetes; microvascular complications; microRNA; serum; biomarkers

2型糖尿病(type 2 diabetes，T2DM)發病機制目前尚不完全清楚。隨著患病時間延長，T2DM患者發生微血管病變(type 2 diabetes microvascular complications，T2DMC)幾率上升，但目前臨床尚缺乏有效診斷T2DMC的血清生物標志物。

微小核糖核酸(microRNA，miRNA)是一類非編碼單鏈小RNA分子，可通過與靶信使RNA 3′端非翻譯序列完全或部分互補結合調控靶基因表達[1]。研究顯示，miRNA與β細胞功能、胰島素分泌和T2DM發生發展密切相關[2]。研究發現，miR-661可參與胰島素合成及調節，與血管內皮細胞功能及內皮細胞向間充質細胞轉化密切相關，提示其可能與T2DM及T2DMC發生發展密切相關[3-4]。人血清/血漿中含有大量穩定表達的miRNA，其對T2DM臨床價值目前尚不清楚，但相關研究較少。……