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中国应用生理学杂志 ›› 2020, Vol. 36 ›› Issue (4): 358-362.doi: 10.12047/j.cjap.5940.2020.077

• 研究论文 • 上一篇    下一篇

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*

田香勤1,2, 谭朝阳1,3, 李娜娜2, 常玉巧2, 张爱梅4, 周颖1, 马克涛1, 司军强   

  1. 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
  • 收稿日期:2019-09-18 修回日期:2020-04-17 发布日期:2020-11-09
  • 通讯作者: Tel: 0993-2057555; E-mail: sijunqiang11@hotmail.com
  • 基金资助:
    *国家自然科学基金资助项目(81560081)

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

TIAN Xiang-qin1,2, TAN Chao-yang1,3, LI Na-na2, CHANG Yu-qiao2, ZHANG Ai-mei4, ZHOU Ying1, MA Ke-tao1, SI Jun-qiang   

  1. 1. Department of Physiology, Medical College of Shihezi University, Shihezi 832002;
    2. Key Open Laboratory for Tissue Regeneration of Henan Province, Xinxiang Medical University, Xinxiang 453003;
    3. Karamay Army Division of Chinese People's Liberation Army, Karamay 834000;
    4. First Affiliated Hospital, Medical College of Shihezi University, Shihezi 832002, China
  • Received:2019-09-18 Revised:2020-04-17 Published:2020-11-09

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。

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

Abstract: 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 Ca2+-activated Cl- current (ICl(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.

Key words: cardiac fibroblasts, calcium-activated chloride channels, ANO1, whole-cell patch clamp, myocardial fibrosis

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