摘要
目的探讨瞬时受体电位通道6(transient receptor potential channel 6,TRPC6)通过调控上皮间质转化(epithelial-mesenchymal transition,EMT)减轻TGF-β1诱导的肾小管上皮细胞(tubular epithelial cells,TEC)损伤的分子机制。方法以TRPC6基因敲除型(TRPC6 knockout,TRPC6^(-/-))和野生型(wild type,WT)小鼠为实验对象,分离培养原代TEC,采用光学显微镜观察并记录细胞生长形态学变化;采用免疫荧光技术检测TEC中β-catenin的表达情况;采用Western blot技术检测TEC中TRPC6、EMT分子标记和GSK-3β/β-catenin通路相关蛋白的表达。结果WT小鼠原代TEC经TGF-β1刺激后,TRPC6表达升高,EMT发生,GSK-3β/β-catenin信号通路被激活。TRPC6基因敲除后,TEC中EMT被抑制,同时GSK-3β/β-catenin信号通路被下调。结论敲除TRPC6基因可能通过GSK-3β/β-catenin信号通路抑制EMT,减轻TGF-β1诱发的TEC损伤。
Objective To investigate the mechanism of transient receptor potential channel 6(TRPC6)alleviating tubular epithelial cell(TEC)injury induced by TGF-β1 through regulation of epithelial-mesenchymal transition(EMT).Methods Primary TEC was isolated and cultured in TRPC6 knockout(TRPC6^(-/-))and wild type(WT)mice.Then cell morphological changes were observed by optical microscope.While the expression ofβ-catenin was detected by immunofluorescence,the expression of TRPC6,EMT molecular markers and GSK-3β/β-catenin related proteins were detect by Western blotting.Results The expression of TRPC6 was significantly increased,along with the appearance of EMT and the activation of GSK-3β/β-catenin signaling pathway in TEC treated with TGF-β1.After deletion of TRPC6,EMT was inhibited,and GSK-3β/β-catenin signaling pathway was downregulated.Conclusion TRPC6 knockout may inhibit EMT through GSK-3β/β-catenin signaling pathway to alleviate TEC injury induced by TGF-β1.
作者
张彦红
韩永明
陈泽斌
廖燕宏
Zhang Yanhong;Han Yongming;Chen Zebin(Department of Anatomy,College of Basic Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China;Department of Anatomy,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China;Key Laboratory of Neurological Diseases of Ministry of Education,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)
出处
《华中科技大学学报(医学版)》
CAS
CSCD
北大核心
2022年第4期443-448,共6页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.31171087)。
关键词
瞬时受体电位通道6
上皮间质转化
肾小管上皮细胞
转化生长因子-β1
transient receptor potential channel 6
epithelial-mesenchymal transition
tubular epithelial cell
transforming growth factor-β1