PTEN对糖尿病肾病肾间质纤维化的作用及其机制  

Effect and mechanism of PTEN on renal interstitial fibrosis in diabetic nephropathy

作　　者：刘兴梅[1] 沈燕[2] 班晓霞 何玉[1] 李红梅[1] 何晓兰 张华[1] Liu Xing-Mei;Shen Yan;Ban Xiao-Xia;He Yu;Li Hong-Mei;He Xiao-Lan;Zhang Hua(Department of Clinical Laboratory,Guizhou Provincial People's Hospital,Guiyang 550002,China;Department of Nephrology,Guizhou Provincial People's Hospital,Guiyang 550002,China;Department of Pathology,Guizhou Provincial People's Hospital,Guiyang 550002,China)

机构地区：[1]贵州省人民医院检验科,贵阳550002 [2]贵州省人民医院肾内科,贵阳550002 [3]贵州省人民医院病理科,贵阳550002

出　　处：《解放军医学杂志》2022年第5期435-441,共7页Medical Journal of Chinese People's Liberation Army

基　　金：贵州省科技计划项目[黔科合基础(2020)1Y305]。

摘　　要：目的探讨同源性磷酸酶-张力蛋白(PTEN)对糖尿病肾病肾间质纤维化的作用及其可能机制。方法将体外培养的大鼠近端肾小管上皮(NRK52E)细胞分为正常糖(NG)组、高糖(HG)组及高渗(MA)组,采用Western blotting检测各组PTEN、LC3、P62、Ⅰ型胶原和Ⅲ型胶原的表达水平。另将NRK52E细胞分为正常糖对照(NG)组、高糖对照(HG)组和高糖过表达PTEN(HG+pPTEN)组,采用激光共聚焦显微镜观察PTEN对自噬的影响,并在荧光显微镜下观察PTEN对胶原表达的影响。结果Western blotting检测结果显示,与NG组比较,HG组PTEN、LC3表达水平明显降低,而P62、Ⅰ型胶原和Ⅲ型胶原表达水平明显升高,差异均有统计学意义(P<0.01),而MA组PTEN、LC3、P62、Ⅰ型胶原和Ⅲ型胶原表达水平无明显变化。激光共聚焦显微镜观察结果显示,HG组红色斑点和黄色斑点数减少,表明存在明显的自噬抑制,而其余两组未见明显自噬抑制。细胞免疫荧光染色结果显示,HG组PTEN红色荧光减弱、表达降低,胶原荧光染色增强、表达升高,而其余两组未见明显变化。结论高糖培养的肾小管上皮细胞中PTEN表达降低,自噬被抑制,进而可加重肾纤维化;过表达PTEN后,可恢复自噬并缓解肾纤维化病变的进展。Objective To explore the effect and mechanism of phosphatase and tensin homolog(PTEN)on renal interstitial fibrosis in diabetic nephropathy.Methods We culture NRK52E cells in vitro,cells were randomly assigned to three groups:normal glucose group(NG),high glucose group(HG)and mannitol group(MA),Western blotting was used to detect the protein levels of PTEN,LC3,P62,collagen-ⅠandⅢin vitro.To detect the effect of PTEN on autophagy and collagen,the NRK52E were randomly assigned to three groups:NG group,HG group and HG+pPTEN group,the autophagy changes were observed by laser confocal microscopy,and the changes of collagen were observed by fluorescence microscopy.Results Western blotting in vivo showed that,compared with NG group,the expression levels of PTEN and LC3 were decreased,but of P62,collagen-ⅠandⅢobviously increased in HG group(P<0.01),while no obvious change of the expression of PTEN and LC3,P62,collagen-ⅠandⅢin MA group.The confocal microscope showed that,the number of red spots and yellow spots were decreased and significant autophagy inhibition in HG group,there were no significant autophagy inhibition in other two groups.The cellular immunofluorescence staining showed that,the red fluorescence of PTEN and protein expression were decreased,the collagen fluorescence staining and protein expression were increased in HG group,there were no significant change in other two groups.Conclusions In NRK52E cells under high glucose,PTEN expression reduced significantly,autophagy was inhibited,and then aggravate renal fibrosis.Overexpression of PTEN can restore autophagy and alleviate the progression of renal interstitial fibrosis.

关 键 词：糖尿病肾病 同源性磷酸酶-张力蛋白 自噬 肾间质纤维化 

分 类 号：R363.2[医药卫生—病理学] R587.2[医药卫生—基础医学]