基于WNT/β-catenin信号通路研究DMBT1调控慢性肾小球肾炎发生发展的作用机制  

A study on mechanisms of DMBT1’s regulation of occurrence and progression of chronic glomerulonephritis based on WNT signaling pathway

作　　者：卢作奎 许为佳 史秀岩[1] LU Zuo-kui;XU Wei-Jia;SHI Xou-yan(Department of Nephropathy,Hubei Province Shiyan Taihe Hospital,Shiyan 442000,China)

机构地区：[1]湖北省十堰市太和医院肾内科,十堰442000

出　　处：《临床肾脏病杂志》2020年第4期297-302,共6页Journal Of Clinical Nephrology

基　　金：2013年湖北省教育厅科研项目(B2013117)。

摘　　要：目的研究DMBT1在人慢性肾小球肾炎(chronic glomerulonephritis,CGN)中相对表达及甲基化水平,分析DMBT1与WNT/β-catenin信号通路在CGN发生发展中的关联。方法收集30例正常人肾组织(肾癌手术中切除的癌旁正常肾组织)、86例CGN患者肾组织作为研究对象,通过实时定量PCR和免疫蛋白印迹实验验证DMBT1在CGN肾组织中的表达,采用甲基化PCR分析DMBT1在CGN患者中甲基化水平。选取人肾小球足细胞分为两组:DMBT1转染组(进行DMBT1基因过表达)和对照组,采用细胞划痕实验研究两组细胞迁移情况;通过免疫蛋白印迹实验验证DMBT1对WNT/β-catenin信号通路的影响。结果与正常肾组织相比,DMBT1在CGN患者肾组织中表达下调(P<0.01);甲基化PCR结果显示CGN患者的DMBT1甲基化程度明显增高;DMBT1基因过表达能抑制细胞迁移和侵袭(P<0.01);过表达DMBT1后,WNT/β-catenin信号通路及其下游细胞因子明显受到抑制。结论DMBT1在CGN中下调,能作为重要的调控因子参与CGN的发生发展。Objective To study the relative expression and methylation level of DMBT1 in human chronic glomerulonephritis(GGN),and to analyze the correlation of DMBT1 with the WNT/β-catenin signaling pathway in occurrence and progression of GGN.Methods The specimens of normal human kidney tissue(normal kidney tissue near the cancer tissue excised during kidney cancer surgery)from 30 cases of normal people,and damaged kidney tissue from 86 cases of GGN patients,were collected.We examined the expression of DMBT1 in the kidney tissue with GGN through real time semi-quantitative polymerase chain reaction(qRT-PCR)and western blot,and analyzed the methylation level of DMBT1 gene in the patients with GGN by methylation polymerase chain peaction(MSP).To detect the migration function of DMBT1,we selected human podocytes(HPC)and divided them into DMBT1 transfection group(to conduct over-expression of NMBT1 gene)and control group.Wound-healing assay was used to investigate migration of the cells in the two groups.Western blot was applied to explore the effect of DMBT1 on the WNT/β-catenin signaling pathway.Results DMBT1 expression was down-regulated in CGN compared with normal kidney tissues(P<0.01).Methylation PCR showed that the methylation level of DMBT1 gene was increased significantly in the GGN patients,over-expression of DMBT1 gene could inhibit cell migration and invasion(P<0.01),and after over-expression of DMBT1 gene the WNT/β-catenin signaling pathway and its downstream cytokines were suppressed obviously.Conclusions As a key regulatory factor,DMBT1 is involved in occurrence and progression of GGN by means of down-regulated methylation of DMBT1 gene.

关 键 词：慢性肾小球肾炎 DMBT1 基因表达 甲基化 WNT/Β-CATENIN信号通路 

分 类 号：R692.31[医药卫生—泌尿科学]