循环miRNA-93在DHEA多囊卵巢综合征大鼠模型中的表达及其意义

The study on the expression and diagnostic value of circulating miRNA-93 in a DHEA rat PCOS mode

目的:通过检测多囊卵巢综合征(PCOS)大鼠血清与卵巢组织中miRNA-93及血管内皮生长因子A(VEGFA)的表达水平,探讨循环miRNA-93及靶基因VEGFA在PCOS发生发展过程中的作用机理与诊断价值。方法:选取3~4周龄雌性(SD)大鼠,随机分为实验组和对照组,每组20只。实验组每日颈背部皮下注射脱氢表雄酮(DHEA),按6mg/100g剂量溶于0.2ml无菌注射用油;对照组每日颈背部皮下注射0.2ml无菌注射用油剂。28天后观察两组大鼠的动情周期、卵巢形态学、性激素、体重、卵巢体积、OGTT血糖、空腹胰岛素、胰岛素抵抗指数(HOMA-IR)及血脂等代谢相关指标的变化。RT-PCR法检测大鼠血清和卵巢组织中miRNA-93表达,ELISA法检测血清中VEGFA表达,IHC、Western blot法分析卵巢组织中VEGFA蛋白表达量。结果:实验组大鼠失去规律的动情周期,卵巢体积增大,呈多囊样变,血清FSH、T、LH升高,糖耐量异常,空腹胰岛素及胰岛素抵抗指数明显高于对照组,血清胆固醇、甘油三酯高于对照组,PCOS大鼠模型构建成功。RT-PCR、ELISA、Western blot、IHC等结果显示,miRNA-93及靶基因VEGFA在PCOS大鼠模型的血清、卵巢组织中表达一致且显著升高(P<0.05)。结论:miRNA-93在PCOS大鼠模型血清和卵巢组织中呈高表达,可能通过调控其靶基因VEGFA在PCOS发生发展中起重要作用,在PCOS诊断中循环miRNA-93可能作为一种新型非侵入性的诊断标记物。

Objective： Through detecting the expressions of miRNA-93 and vascular endothelial growth factor A（ VEGFA） in serum and ovary tissue of rats with PCOS,to explore the mechanism and diagnostic value of circulating miRNA-93 and target gene VEGFA in the development and progression of PCOS.Methods： 3 ~ 4 week old female（ SD） rats were randomly average divided into control group and experimental group. The experimental group received DHEA 6 mg/100 g weight and the control group received 0.2 ml injection oil respectively by subcutaneous injection on neck and back daily.After 28 days,the estrous cycle,ovarian morphologysex hormone,body weight,ovarian volume,OGTT blood glucose,fasting insulin,insulin（ HOMA-IR） changes and lipid metabolism of the two groups of rats were tested. The miRNA-93 in serum and ovarian tissue was detected by real-time quantitative PCR（ RT-PCR）,and the VEGFA in serum was detected by enzyme linked immunosorbent assay（ ELISA）.The VEGFA in ovarian tissue was detected by immunohistochemistry（ IHC） and Western blot. Results： Themodel rat groups were the regular estrous cycle lost,ovarian volume enlarged and polycystic,serum FSH,T and LH increased,glucose tolerance abnormal.The fasting insulin and insulin resistance index was significantly higher than the control group.Meanwhile the serum cholesterol and triglyceride were higher than the control group. The model of PCOS rats was successfully constructed.RT-PCR,ELISA,Western blot,IHC and other assay results showed that the expressions of miRNA-93 and target gene VEGFA in the serum and ovary tissues of PCOS rats increased significantly（ P〈0.05）.Conclusions： The miRNA-93 in serum and ovary tissues of PCOS rats was highly expressed and may play an important role in the development of PCOS by regulating its target gene VEGFA.Circulating miRNA-93 may represent a novel non-invasive diagnostic for PCOS.