实验动物与比较医学 ›› 2020, Vol. 40 ›› Issue (4): 270-.DOI: 10.3969/j.issn.1674-5817.2020.04.002

• 论著 • 上一篇    下一篇

小泛素样修饰物蛋白酶3调控小鼠肺泡Ⅱ型上皮细胞自噬

朱慧琴1,邹嫣琼2,杨   洁2,易   静1,杨   洁1   

  1. 1. 上海交通大学医学院生物化学与分子细胞生物学系,上海 200025;
    2. 上海交通大学医学院基础医学公共技术平台,上海 200025
  • 收稿日期:2019-12-31 出版日期:2020-08-25 发布日期:2020-11-23
  • 作者简介:朱慧琴(1980—), 女, 实验师, 研究方向: 蛋白质翻译后修饰。E-mail: zhuhuiqin21@yeah.net
  • 基金资助:
    国家自然科学基金(31771522); 上海市自然科学基金(16ZR1418400)

Small Ubiquitin-like Modifier Specific Protease 3 Regulates Autophagy in Mouse Alveolar TypeⅡ Epithelial Cells

ZHU Huiqin1, ZOU Yanqiong2, YANG Jie2, YI Jing1, YANG Jie1   

  1. 1. Department of Biochemistry and Molecular Cell Biology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China;  2. Core Facility of Basic Medical Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Received:2019-12-31 Online:2020-08-25 Published:2020-11-23

摘要:

目的   探讨小泛素样修饰物(small ubiquitin-like modifier,SUMO)特异性蛋白酶3(SUMO-specific protease 3,SENP3)对小鼠肺组织细胞自噬的调控作用及分子机制。方法    应用免疫荧光技术检测SENP3在肺组织细胞中的定位;对SENP3基因野生型C57BL/6J小鼠(SENP3+/+)和SENP3基因敲除杂合子小鼠(SENP3+/-)进行饥饿处理,诱导细胞自噬;应用免疫印迹法评估细胞自噬水平;应用电子显微镜观察和免疫荧光技术分析发生自噬的细胞类型;应用免疫共沉淀方法检测自噬相关分子卷曲螺旋状Myosin样Bcl-2相互作用蛋白1(coiled-coil myosin-like Bcl-2-interacting protein 1,BECN1)的SUMO化修饰。结果    SENP3在肺泡Ⅱ型上皮细胞中高表达。小鼠经饥饿处理后,肺泡Ⅱ型上皮细胞出现自噬现象,且SENP3+/-小鼠较 SENP3+/+小鼠更明显。同时,肺组织样品中BECN1的SUMO2/3化修饰被SENP3去除。结论    SENP3抑制饥饿应激时小鼠肺泡Ⅱ型上皮细胞的自噬,并可对细胞自噬程度进行精细调控。


关键词: 小泛素样修饰物蛋白酶3, 自噬, 肺泡Ⅱ型上皮细胞, 小鼠

Abstract:  Objective     To explore in vivo the role and molecular mechanism of small ubiquitin-like modifier (SUMO) specific protease 3 (SENP3) in regulating autophagy in mouse lung. Methods   Immunofluorescence was performed to detect SENP3 localization in lung tissues. SENP3+/-       and SENP3+/+ mice were starved to induce autophagy. Immunoblotting was used to evaluate the overall autophagy level. Electron microscopy and immunofluorescence were used to detect autophagy, and to identify which type(s) of cells undergoing autophagy in mouse lung. Co-immunoprecipitation was used to detect the SUMOylation modification of autophagy-related molecule coiled-coil myosin-like Bcl-2-interacting protein 1 (BECN1). Results     SENP3 was highly expressed in alveolar typeⅡ epithelial cells in lung tissues. After starvation, the mouse alveolar type Ⅱ epithelial cells underwent autophagy, and SENP3+/- mice was more pronounced than SENP3+/+ mice. SUMO2/3 modification of BECN1 in lung tissue samples was removed by SENP3. Conclusion     SENP3 plays a role on regulating the degree of autophagy of mouse alveolar typeⅡ epithelial cells during starvation in fine-tuning manner.

Key words: Small ubiquitin-like modifier specific protease 3, Autophagy, Alveolar typeⅡ epithelial cells, Mice

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