Glycogen synthase kinase‑3β is required for epithelial‑mesenchymal transition and barrier dysfunction in mouse podocytes under high glucose conditions.
Glycogen synthase kinase‑3β is required for epithelial‑mesenchymal transition and barrier dysfunction in mouse podocytes under high glucose conditions.
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高糖条件下小鼠足细胞上皮间质转化和屏障功能障碍需要糖原合酶激酶 3 beta
DOI：
10.3892/mmr.2016.5786
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发表时间：
2016-11
影响因子：
3.4
通讯作者：
Liu Z
中科院分区：
文献类型：
--
作者：
Guo J;Yang L;Qiao Y;Liu Z
上皮-间充质转化(EMT)在糖尿病肾病(DN)中起重要作用。足细胞是一种特殊的上皮细胞，构成肾小球滤过屏障的主要组成部分。足细胞损伤被认为是糖尿病肾病相关蛋白尿的主要机制。本研究旨在探讨糖原合成酶-3β在高糖环境下小鼠足细胞内膜转化和屏障功能障碍中的作用。成熟和分化的足细胞分别用正常葡萄糖(NG)、HG或NG+甘露醇处理。在NG或HG条件下，用针对GSK-3β的小干扰RNA或干扰的小干扰RNA或用GSK-3β抑制剂氯化锂处理足细胞。免疫印迹分析和免疫荧光染色分别检测足细胞上皮细胞标志物newitin和podocin和肌成纤维细胞标志物α-平滑肌肌动蛋白和纤维连接蛋白的表达水平。通过白蛋白流入评价单层屏障功能。定量检测GSK-3β的磷酸化水平和活性水平。观察到HG促进足细胞内的EMT，这是由于Podocin和neparin的表达水平增加，而α-SMA和FN的表达水平降低。HG还可引起足细胞屏障功能障碍，增加足细胞总GSK-3β、Try216磷酸化GSK-3β的表达水平和GSK-3β活性。GSK-3βsiRNA或氯化锂处理可逆转HG诱导的足细胞EMT和屏障功能障碍。综上所述，本研究确定GSK-3β是HG条件下足细胞内膜转化和屏障功能障碍所必需的；因此，GSK-3β可能成为治疗糖尿病肾病的新靶点。
Epithelial-mesenchymal transition (EMT) is important for diabetic nephropathy (DN). Podocytes are specialized epithelial cells, which form a major component of the glomerular filtration barrier. Podocyte damage has been suggested to be the primary mechanism behind the albuminuria associated with DN. The present study aimed to determine the function of glycogen synthase kinase (GSK)-3β in EMT and barrier dysfunction of mouse podocytes exposed to high glucose (HG) conditions. Matured and differentiated podocytes were treated with normal glucose (NG), HG or NG + mannitol. Podocytes were also transfected with a small interfering RNA (siRNA) against GSK-3β or a scrambled siRNA, or were treated with lithium chloride (LiCl), a GSK-3β inhibitor, under NG or HG conditions. The expression levels of the epithelial cell markers, nephrin and podocin, and the myofibroblast cell markers, α-smooth muscle actin (SMA) and fibronectin, in podocytes by western blot analysis and immunofluorescence staining, respectively. The monolayer barrier function was assessed by albumin inflow. The phosphorylation and activity levels of GSK-3β were also quantified. It was observed that HG promotes EMT in podocytes, due to the increased levels of podocin and nephrin expression and the reduced α-SMA and fibronectin expression levels. HG also induced barrier dysfunction and increased the expression level of total GSK-3β, Try216-phosphorylated-GSK-3β and the GSK-3β activity in podocytes. Transfection of GSK-3β siRNA or treatment with LiCl reversed the HG-induced EMT and barrier dysfunction in podocytes. In conclusion, the present study determined that GSK-3β is required for EMT and barrier dysfunction in podocytes under HG conditions; therefore, GSK-3β may be a novel target for the treatment of DN.
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影响因子：
15.9
作者：
Soda, Keita;Balkin, Daniel M.;Ishibe, Shuta
通讯作者：
Ishibe, Shuta
影响因子：
8.7
作者：
Hautmann, MB;Adam, PJ;Owens, GK
通讯作者：
Owens, GK
影响因子：
6
作者：
Li, Yingjian;Kang, Young Sun;Liu, Youhua
通讯作者：
Liu, Youhua
影响因子：
13.6
作者：
Lin, Chun-Liang;Wang, Jeng-Yi;Wang, Feng-Sheng
通讯作者：
Wang, Feng-Sheng
DOI：
10.1152/ajprenal.00145.2003
发表时间：
2003-12-01
影响因子：
4.2
作者：
Sharma, K;Deelman, L;Hajnoczky, G
通讯作者：
Hajnoczky, G