吡格列酮调控β-连环蛋白表达延缓糖尿病大鼠肾脏纤维化的组织和细胞实验研究  

Experimental study of Pioglitazone regulates β-catenin expression and delaying renal fibrosis tisssue and cells in diabetic rats

作　　者：刘忠强 屈玲玲 邹琴 田平平 卢雨微 张帆 张国忠 郭兵 石明隽 LIU Zhongqiang;QU Lingling;ZOU Qin(Department of Pathophysiology,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州医科大学病理生理教研室,贵阳550025 [2]贵州省重大疾病发病机制研究及药物防治特色重点实验室

出　　处：《中国糖尿病杂志》2020年第7期529-536,共8页Chinese Journal of Diabetes

基　　金：国家自然科学基金(81860135);贵州省科学术厅学术新苗项目(黔科合[2017]5718-20);贵州医科大学博士启动基金(YJ[2017]23)。

摘　　要：目的在组织和细胞水平研究吡格列酮通过调控Wnt/β-连环蛋白(β-catenin)通路延缓糖尿病大鼠肾脏纤维化的发生发展。方法18只SD大鼠随机分为正常对照(NC)组、DKD组和吡格列酮治疗组(PIO),每组各6只。HE、Masson染色观察肾组织形态学变化。大鼠肾小管上皮细胞(NRK-52E)分为正常糖组(Con)、高糖组(HG)、吡格列酮处理组(HG+PIO)、HG+空载组(HG+E)和HG+PPARγ过表达组(HG+PPARγ)。免疫细胞荧光观察细胞中β-catenin表达;Western blot法检测大鼠肾组织和NRK-52E细胞相关蛋白表达变化。结果与NC组比较,DKD组PPARγ表达降低(P<0.05),磷酸化糖原合成酶激酶3β(p-GSK3β)、β-catenin、细胞核β-catenin、Ⅳ型胶原(CollagenⅣ)、Ⅲ型胶原(CollagenⅢ)和Ⅰ型胶原(CollagenⅠ)蛋白表达升高(P<0.05),与DKD组比较,PIO组PPARγ表达升高(P<0.05),p-GSK3β、β-catenin、细胞核β-catenin、CollagenⅣ、CollagenⅢ和CollagenⅠ表达降低(P<0.05)。与Con组比较,HG组PPARγ降低(P<0.05),p-GSK3β、β-catenin、细胞核β-catenin、CollagenⅣ、CollagenⅢ和CollagenⅠ蛋白表达升高(P<0.05),与HG组比较,HG+PIO组PPARγ表达升高,p-GSK3β、β-catenin、细胞核β-catenin、CollagenⅣ、CollagenⅢ和CollagenⅠ表达降低(P<0.05)。与HG组比较,HG+PPARγ组PPARγ表达升高(P<0.05),p-GSK3β、β-catenin、CollagenⅣ和CollagenⅢ表达降低(P<0.05)。结论吡格列酮可上调PPARγ表达,下调β-catenin表达,抑制Wnt/β-catenin通路活化,减少ECM沉积,延缓糖尿病肾脏纤维化的发生发展。Objective To investigate whether Pioglitazone could delay the onset and development of renal fibrosis in diabetic rats by regulating the Wnt/β-catenin pathway from in vivo and in vitro studies.Methods A total of 18 SD rats were randomly divided into three groups:normal control group(NC,n=6),rats with diabetic kidney disease(DKD,n=6),and Pioglitazone treatment group(PIO,n=6).HE and Masson staining were used to evaluate the morphological changes of kidney tissues.NRK-52E cells were divided into five groups:normal glucose control group(Con),high glucose group(HG),high glucose and Pioglitazone treatment group(HG+PIO),high glucose and no-load vector group(HG+E)and high glucose and PPARγover-expression group(HG+PPARγ).β-catenin expression was observed by immunofluorescence,Western blot was used to detect the expression of rat kidney tissues related and NRK-52E cellrelated proteins.Results Compared with NC group,PPARγdecreased(P<0.05),while the expression of p-GSK3β,β-catenin,nuclearβ-catenin,CollagenⅣ,CollagenⅢand CollagenⅠincreased in DKD group(P<0.05).Compared with DKD group,PPARγincreased(P<0.05),while the expressions of p-GSK3β,β-catenin,nuclearβ-catenin,CollagenⅣ,CollagenⅢand CollagenⅠproteins decreased in PIO group(P<0.05).Compared with Con group,PPARγdecreased,while the expression of p-GSK3β,β-catenin,nuclearβ-catenin,CollagenⅣ,CollagenⅢand CollagenⅠincreased in HG group(P<0.05).Compared with HG group,PPARγincreased(P<0.05),while the expressions of p-GSK3β,β-catenin,CollagenⅣand CollagenⅢdecreased in HG+PIO group(P<0.05).Compared with HG group,PPARγincreased(P<0.05),while the expressions of p-GSK3β,β-catenin,CollagenⅣand CollagenⅢdecreased in HG+PPARγgroup(P<0.05).Conclusion Pioglitazone could up-regulate the expression of PPARγ,down-regulateβ-catenin expression,thereby inhibit Wnt/β-catenin pathway activation and reduce extracellular matrix deposition,and further more,delay the onset and development of diabetic renal fibrosis.

关 键 词：SD大鼠 糖尿病肾脏疾病 吡格列酮 过氧化物酶体增殖物激活受体Γ Β-连环蛋白 

分 类 号：R965[医药卫生—药理学]