利用肾系膜细胞3D培养体系探讨高糖环境下miR-382与肾纤维化的相关性

doi:10.13418/j.issn.1001-165x.2020.01.010

中国临床解剖学杂志 ›› 2020, Vol. 38 ›› Issue (1): 45-50.doi: 10.13418/j.issn.1001-165x.2020.01.010

利用肾系膜细胞3D培养体系探讨高糖环境下miR-382与肾纤维化的相关性

李想¹，　杨志峰²，　王国保¹

1.南方医科大学附属南方医院肾内科，广州 510515；　2.郑州大学第五附属医院全科医学科，郑州 450052

收稿日期:2019-10-29 出版日期:2020-01-25 发布日期:2020-02-18
通讯作者: 王国保，教授，博士生导师，主任医师，E-mail：wgbandyl@medmail.com.cn
作者简介:李想（1994-）男，江西九江人，硕士研究生，研究方向：IgA肾病，E-mail：risou1222@163.com；共同第一作者：杨志峰（1990-），男，河南漯河人，医师，研究方向：糖尿病肾病，E-mail：1427622671@qq.com
基金资助:
国家自然科学基金面上项目（81670669）

Studying the effect of miR-382 on renal fibrosis in high glucose based on 3D culture system of renal mesangial cells

LI Xiang¹, YANG Zhi-feng², WANG Guo-bao¹

1.Department of Nephrology, Nanfang Hospital, Southern Medical University, Guangzhou510515, Guangdong Province, China; 2.Department of General Practice, The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan province, China

Received:2019-10-29 Online:2020-01-25 Published:2020-02-18

摘要/Abstract

摘要： 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。

关键词: , LBL；　3D；　高糖；　miR-382；　肾纤维化

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.

Key words: LBL;　3D;　High glucose;　miR-382, 　Renal fibrosis

中图分类号:

R692.6

李想, 杨志峰, 王国保. 利用肾系膜细胞3D培养体系探讨高糖环境下miR-382与肾纤维化的相关性[J]. 中国临床解剖学杂志, 2020, 38(1): 45-50.

LI Xiang, YANG Zhi-feng, WANG Guo-bao. Studying the effect of miR-382 on renal fibrosis in high glucose based on 3D culture system of renal mesangial cells[J]. Chinese Journal of Clinical Anatomy, 2020, 38(1): 45-50.

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利用肾系膜细胞3D培养体系探讨高糖环境下miR-382与肾纤维化的相关性

Studying the effect of miR-382 on renal fibrosis in high glucose based on 3D culture system of renal mesangial cells

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