目的 采用LBL技术构建肾系膜细胞3D细胞球，研究miR-382在糖尿病肾病系膜病变及肾纤维化中的作用。 方法 采用LBL技术构建肾系膜细胞3D细胞球，研究不同葡萄糖浓度对肾系膜细胞的影响及miR-382在其中的作用。实验组分为3组（葡萄糖浓度分别设为5.6 mmol/L、25 mmol/L、40 mmol/L），采用qRT-PCR、Western blotting、免疫荧光等手段检测各组细胞中转化生长因子（TGF-β1）、纤连蛋白（Fn）的表达，miR-382及其可能的靶基因第10号染色体缺失的磷酸酶和张力蛋白同源基因（PTEN）的水平。 结果 高糖可使系膜细胞miR-382表达上调，分泌PTEN减少，TGF-β1、Fn表达显著增加。 结论 miR-382可能通过负向调控PTEN的表达，参与糖尿病肾病早期系膜病变及肾纤维化过程。
Abstract
Objective To explore the effect of miR-382 in diabetic nephropathy and renal fibrosis by constructing 3D renal mesangial cells spheroid with LBL. Methods LBL was utilized to construct 3D renal mesangial cells spheroid for studying the effect of different concentrations of glucose on renal mesangial cells and the effect of miR-382 on it. The experimental groups were divided into three groups according to the concentrations of glucose (5.6 mmol/L, 25 mmol/L, 40 mmol/L). qRT-PCR, Western blotting and immunofluorescence were used to detect the expression of TGF-β1, Fn, miR-382 and the level of its possible target gene the 10th chromosome deficiency phosphatase and tension of homologous gene (PTEN). Results High glucose upregulated the expression of miR-382, downregulated secretion of PTEN, and significantly increased the expression of TGF-β1 and Fn. Conclusions miR-382 may be involved in the pathology lesion of mesangial cells in the early state of diabetic nephropathy and renal fibrosis via negatively regulating the expression of PTEN.
关键词
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LBL； 3D； 高糖； miR-382； 肾纤维化
Key words
LBL; 3D; High glucose; miR-382 /
Renal fibrosis
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基金
国家自然科学基金面上项目（81670669）
京ICP备08002354号-3
