WISP-1对肾纤维化大鼠肾小管上皮细胞纤维化的影响  

Effect of WISP-1 on fibrosis of renal tubular epithelial cell in rats with renal fibrosis

作　　者：杨二梅 夏薇青 宋培[1] 严佳 YANG Ermei;XIA Weiqing;SONG Pei;YAN Jia(Department of Nephrology,the Third Hospital of Shijiazhuang,Shijiazhuang 050011,China)

机构地区：[1]石家庄市第三医院肾内科,河北石家庄050011

出　　处：《基础医学与临床》2025年第4期471-477,共7页Basic and Clinical Medicine

基　　金：2021年河北省医学科学研究重点课题计划(20210528)。

摘　　要：目的探究Wnt分泌蛋白1(WISP-1)对肾纤维化大鼠肾小管上皮细胞纤维化的影响及机制。方法选取24只成年大鼠及大鼠肾小管上皮细胞系(NEK-52E),分别进行体内及体外实验。大鼠随机分为对照组及研究组,采用单侧输尿管闭塞手术构建体内肾纤维化模型,研究组使用WISP-1/CCN4亲和纯化的单克隆抗体注射阻断肾脏WISP-1表达;细胞系培养后诱导纤维化,分别给予WISP-1过表达及抑制处理,采用组织学和免疫组织化学检测肾脏大鼠病理学变化,使用Western法及RT-qPCR法检测纤维化相关指标。结果WISP-1过表达胶原蛋白Ⅰ(ColⅠ)、纤连蛋白(FN)、转化生长因子-β1(TGF-β1)蛋白及mRNA高于对照,WISP-1抑制ColⅠ、FN、TGF-β1蛋白及mRNA低于对照(P<0.05);WISP-1阻断给药减弱肾纤维化其病理变化。研究组ColⅠ、FN、α平滑肌肌动蛋白(α-SMA)、TGF-β1蛋白及mRNA、LC3、Beclin-1蛋白低于对照组,泛素结合蛋白p62(SQSTM1/p62)高于对照组(P<0.05)。WISP-1抑制剂LC3、Beclin-1的表达低于对照组,SQSTM1/p62的表达高于对照组,WISP-1过表达组LC3和Beclin-1的表达高于对照组,SQSTM1/p62的表达低于对照组。结论WISP-1过表达促进大鼠肾小管上皮细胞纤维化,增强TGF-β1介导的自噬可能是肾纤维化的机制之一。Objective To investigate the effect and mechanism of WISP-1 on renal tubular epithelial cell fibrosis in rats with renal fibrosis.Methods Twenty-four adult rats and renal tubular epithelial cell line(NEK-52E)were selected for in vivo and in vitro experiments.The rats were randomly divided into control group and study group,and the in vivo renal fibrosis model was established.The expression of WISP-1 in the study group was blocked.The cell lines were induced to fibrosis followed by WISP-1 over-expression or inhibition.The related indicators of fibrosis were observed.Results Protein and mRNA level of CollagenⅠ(ColⅠ),fibronectin(FN),transforming growth factor-β1(TGF-β1)in WISP-1 over-expression group were all higher than those in control group.The protein and mRNA level of ColⅠ,FN and TGF-β1 in the WISP-1 inhibition group were lower than those in the control group(P<0.05).WISP-1 blockade attenuated the pathological changes of renal fibrosis.The protein and mRNA levels of ColⅠ,FN,α-smooth muscle actin(α-SMA),TGF-β1,LC3 and Beclin-1 in the study group were lower than those in the control group,and the level of ubiquitin binding protein p62(SQSTM1/p62)was higher than that in the control group(P<0.05).The expression of LC3 and Beclin-1 in WISP-1 inhibitor group was lower than that in the control group and the expression of SQSTM1/p62 was higher than that in the control group.The expression of LC3 and Beclin-1 in the WISP-1 over-expression group was higher than that in the control group,and the expression of SQSTM1/p62 was lower than that in the control group.Conclusions Overexpression of WISP-1 promotes the fibrosis of rat renal tubular epithelial cells,and enhanced TGF-β1-mediated autophagy might be a underlying mechanism to mediate renal fibrosis.

关 键 词：Wnt分泌蛋白1 慢性肾脏病 肾纤维化 作用机制 

分 类 号：R363[医药卫生—病理学]