激酶锚定蛋白12/蛋白原转化酶枯草杆菌蛋白酶/kexin型6信号通路对糖尿病肾脏疾病小鼠足细胞线粒体自噬保护作用的研究  

Protective effect of a kinase anchor protein 12 on podocyte mitochondrial autophagy in diabetic kidney disease

作　　者：赖轻舟 吴小琴[1] 龚玉萍[1] LAI Qingzhou;WU Xiaoqin;GONG Yuping(Department of Endocrinology,Pingxiang People’s Hospital,Pingxiang 337000,China)

机构地区：[1]萍乡市人民医院内分泌科,337000

出　　处：《中国糖尿病杂志》2024年第10期770-777,共8页Chinese Journal of Diabetes

基　　金：江西省卫生健康委普通科技计划(20187199)。

摘　　要：目的 探讨激酶锚定蛋白 12(AKAP12)/蛋白原转化酶枯草杆菌蛋白酶/kexin型 6(PCSK6)信号通路对DKD小鼠足细胞线料体自噬的保护作用.方法 20 只C57/B6 背景的AKAP12 KO(AKAP12-/-)小鼠和20 只野生型(WT)同窝出生小鼠,随机分为WT小鼠对照组(Ctrl+WT组)、AKAP12-/-小鼠对照组(Ctrl+AKAP12-/-组)、WT小鼠诱导DKD模型组(DKD+WT组)和AKAP12-/-小鼠诱导DKD模型组(DKD+AKAP12-/-组),每组各10 只.从AKAP12-/-小鼠和WT小鼠获得原代足细胞,并暴露于高糖(HG,30 mmol/L)或甘露醇(Mannitol)24 h,分为Mannitol+WT组、HG+WT组、Mannitol+AKAP12-/-、HG+AKAP12-/-组,观察足细胞线粒体分裂和自噬情况.采用sh-PCSK6 转染AKAP12-/-足细胞以敲低PCSK6 表达,将细胞分为Mannitol+阴性对照组(Mannitol+sh-NC组)、Mannitol+PCSK6 敲除载体组(Mannitol+sh-PCSK6 组)、HG+sh-NC组和HG+sh-PCSK6组.采用Mito-Tracker染色分析足细胞中线粒体的形态.Western blot法检测线粒体分裂蛋白(FIS1和DRP1)、线粒体自噬(PINK1 和Parkin)和自噬相关(LC3 和p62)蛋白表达.结果 与HG+WT组比较,HG+AKAP12-/-组足细胞中单个线粒体数量、FIS1、DRP1、PINK1、Parkin、LC3Ⅱ、p62 蛋白表达升高(P<0.05),线粒体平均分支长度降低(P<0.05).与HG+sh-NC组比较,HG+sh-PCSK6组足细胞单个线粒体数量、FIS1、DRP1、PINK1、Parkin和LC3Ⅱ蛋白表达降低(P<0.05 或P<0.01),线粒体平均分支长度升高(P<0.01).结论 AKAP12/PCSK6 信号通路介导了HG环境下足细胞中线粒体分裂过程和线粒体自噬的调节.Objective To investigate the protective effect of AKAP12/PCK6 signaling pathway on podocyte autophagy in diabetic kidney disease(DKD).Methods A total of 40 C57/B6 AKAP12 KO(AKAP12‑/‑)mice and wild‑type(WT)littermates were randomly divided into four groups,with 10 mice in each group:Ctrl+WT group,Ctrl+AKAP12‑/‑group,DKD+WT group and DKD+AKAP12‑/‑group.Primary podocytes were obtained from AKAP12‑/‑mice and WT mice,and exposed to high glucose(HG,30 mmol/L)or mannitol for 24 h to investigate the mitosis and autophagy of podocytes.The primary podocytes were divided into Mannitol+WT,G+WT,Mannitol+AKAP12‑/‑and HG+AKAP12‑/‑.In addition,AKAP12‑/‑podocytes were transfected with sh‑PCSK6 to knockdown the expression of PCK6.Mito‑Tracker staining was used to analyze the morphology of mitochondria in podocytes.The expressions of mitotic proteins(FIS1 and DRP1),mitochondrial autophagy(PINK1 and Parkin)and autophagy‑related proteins(LC3 and p62)were analyzed by Western blot.Results Compared with HG+WT group,the number of single mitochondria and the expression of FIS1,DRP1,PINK1,Parkin,LC3Ⅱ and p62 proteinsincreased,while the average branch length of mitochondria decreased in HG+AKAP12‑/‑ group( P<0. 05).Compared with HG+sh‑NC group,the number of single mitochondria and the expression of FIS1,DRP1,PINK1,Parkin and LC3Ⅱ proteins decreased significantly in HG+sh‑PCSK6 group( P<0. 05 or P<0. 01),while theaverage branch length of mitochondria increased significantly( P<0. 05). Conclusions AKAP12/PCSK6signaling pathway mediates the regulation of mitochondrion division and mitochondrion autophagy inpodocytes under HG environment.

关 键 词：激酶锚定蛋白12 线粒体自噬 蛋白原转化酶枯草杆菌蛋白酶/kexin型6 糖尿病肾脏疾病 足细胞 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]