Abstract:
To investigate the expression level of miR-25-3p in patients with type 2 diabetes mellitus (T2DM) and diabetic nephropathy (DN), and its effect on proliferation, apoptosis and inflammatory response of mesangial cells cultured with high glucose. Blood samples of all clinical subjects were collected for RT-qPCR analysis to detect serum miR-25-3p levels. Human mesangial cells (HMCs) cultured with high glucose were used to construct DN model in vitro. MTT assay, flow cytometry and ELISA were used to evaluate the effects of miR-25-3p on the proliferation, apoptosis, and inflammatory response of DN cell models. Serum miR-25-3p was decreased in both T2DM group and DN group, but more in DN group. Serum miR-25-3p was positively correlated with eGFR and negatively correlated with UAER. The expression of miR-25-3p was reduced in HMCs induced by high glucose. Transfection of miR-25-3p mimic could significantly up-regulate the miR-25-3p level in HMCs. Besides, high glucose culture resulted in abnormal proliferation of HMCs, reduced apoptotic cells, and increased inflammation. The addition of miR-25-3p mimic significantly inhibited cell proliferation and promoted cell apoptosis and reduced the production of inflammatory factors. The abnormal reduction of serum miR-25-3p in DN indicates that it may be a potential biomarker for clinical diagnosis of DN. In in vitro experiments, miR-25-3p was involved in the progression of DN by regulating cell proliferation, apoptosis, and inflammatory response.
摘要:
探讨miR-25-3p在2型糖尿病(T2DM)和糖尿病肾病(DN)患者中的表达水平。以及它对增殖的影响,高糖培养肾小球系膜细胞的凋亡和炎症反应。收集所有临床受试者的血液样品用于RT-qPCR分析以检测血清miR-25-3p水平。采用高糖培养的人肾小球系膜细胞(HMCs)体外构建DN模型。MTT测定,用流式细胞术和ELISA检测miR-25-3p对小鼠增殖的影响,凋亡,和DN细胞模型的炎症反应。血清miR-25-3p在T2DM组和DN组降低,但更多的DN组。血清miR-25-3p与eGFR呈正相关,与UAER呈负相关。高糖诱导的HMC中miR-25-3p的表达降低。转染miR-25-3p模拟物可以显著上调HMC中的miR-25-3p水平。此外,高糖培养导致HMC异常增殖,减少凋亡细胞,增加炎症。添加miR-25-3p模拟物显著抑制细胞增殖,促进细胞凋亡,减少炎症因子的产生。血清miR-25-3p在DN中的异常降低提示其可能是临床诊断DN的潜在生物标志物。在体外实验中,miR-25-3p通过调节细胞增殖参与DN的进展,凋亡,和炎症反应。