钙激活氯通道蛋白ANO1在心肌成纤维细胞分化过程中的表达及其钙激活氯通道电生理特性变化  

Expression of ANO1 during cardiac fibroblasts differentiation and its electrophysiological characteristics as calcium-activated chloride channel protein

作　　者：田香勤 谭朝阳[1,3] 李娜娜 常玉巧 张爱梅 周颖[1] 马克涛 司军强[1] TIAN Xiang-qin;TAN Chao-yang;LI Na-na;CHANG Yu-qiao;ZHANG Ai-mei;ZHOU Ying;MA Ke-tao;SI Jun-qiang(Department of Physiology,Medical College of Shihezi University,Shihezi 832002;Key Open Laboratory for Tissue Regeneration of Henan Province,Xinxiang Medical University,Xinxiang 453003;Karamay Army Division of Chinese People's Liberation Army,Karamay 834000;First Affiliated Hospital,Medical College of Shihezi University,Shihezi 832002,China)

机构地区：[1]石河子大学医学院生理教研室,新疆石河子832000 [2]新乡医学院河南省高等学校组织再生重点开放实验室,河南新乡453003 [3]中国人民解放军克拉玛依军分区,新疆克拉玛依834000 [4]石河子大学医学院第一附属医院,新疆石河子832000

出　　处：《中国应用生理学杂志》2020年第4期358-362,I0002,共6页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助项目(81560081)。

摘　　要：目的:研究钙激活氯通道蛋白ANO1在心肌成纤维细胞(CFs)向成肌纤维细胞(MFs)分化过程中的表达规律及其电生理特性,探索ANO1在心肌纤维化进程中的作用。方法:以刚分离贴壁的大鼠原代CFs及其分化的MFs为研究对象,利用全细胞膜片钳检测钙激活氯通道电流变化,采用免疫荧光标记技术和Western blot检测ANO1、α-SMA和vimentin在CFs和MFs细胞中的表达特征。结果:刚贴壁培养的CFs钙激活氯通道电流密度远高于MFs;ANO1主要表达于CFs和MFs的细胞核周围及细胞核上,少量表达于细胞膜;ANO1在刚贴壁的CFs中表达较弱,而在有分裂增殖趋向的MFs细胞中表达较为明显。结论:ANO1的表达与MFs的分化过程密切相关,提示其可能参与调节心肌纤维化进程。Objective:To investigate the expression and electrophysiological characteristics of calcium-activated chlorine channel anoctamin-1(ANO1)protein during the differentiation of cardiac fibroblasts(CFs)into myofibroblasts(MFs),and to elucidate the role of ANO1 in myocardial fibrosis.Methods:The primary CFs from neonatal rats were isolated and the cells differentiated into MFs by subculture.The Ca 2+-activated Cl-current(I Cl(Ca))in CFs and MFs were measured by whole-cell patch clamp,and the expressions of ANO1,α-smooth muscle actin(α-SMA)and vimentin in CFs and MFs were detected by immunofluorescence assay and Western blot,respectively.Results:The current density in the early adherent CFs was stronger than that in MFs.ANO1 was expressed preferentially within and around the nuclei,and a small amount of ANO1 was expressed on the cell membrane.Moreover,ANO1 expression was weak in the early adherent CFs and displayed stronger expression in the MFs with proliferation tendency.Conclusion:The expression of ANO1 is closely related to the differentiation of MFs and it may be involved in modulation myocardial fibrosis.

关 键 词：心肌成纤维细胞 钙激活氯通道 ANO1 全细胞膜片钳 心肌纤维化 

分 类 号：R331.3[医药卫生—人体生理学]