低氧对裸鼹鼠皮肤成纤维细胞自噬水平及凋亡的影响

doi:10.3969/j.issn.1674-5817.2014.05.013

实验动物与比较医学 ›› 2014, Vol. 34 ›› Issue (5): 405-410.DOI: 10.3969/j.issn.1674-5817.2014.05.013

低氧对裸鼹鼠皮肤成纤维细胞自噬水平及凋亡的影响

赵善民, 林丽芳, 肖邦, 王运慧, 宫晨, 张璐, 余琛琳, 汤球, 孙伟, 崔淑芳

第二军医大学实验动物中心,上海200433

收稿日期:2014-08-06 出版日期:2014-10-25 发布日期:2014-10-25
作者简介:赵善民(1986-), 男, 助教, E-mail:zhaoshanmin2006@163.com
基金资助:
国家自然科学基金项目(No.31402028)、上海市科技发展基金项目(No.12140900200)与(No.13140900400)联合资助

Comparative Effects of Hypoxia on Autophagy and Apoptosis of Naked Mole Rat Fibroblasts

ZHAO Shan-min, LIN Li-fang, XIAO Bang, WANG Yun-hui, GONG Chen, ZHANG Lu, YU Chen-lin, TANG Qiu, SUN Wei, CUI Shu-fang

Laboratory Animal Center of Second Military Medical University, Shanghai 200433, China

Received:2014-08-06 Online:2014-10-25 Published:2014-10-25

摘要/Abstract

摘要： 目的比较裸鼹鼠成纤维细胞常氧与低氧条件下自噬与凋亡水平。方法分别采用低氧(3% O₂)与常氧(20% O₂)培养裸鼹鼠皮肤成纤维细胞,采用Western blotting、RT-PCR等方法检测自噬相关基因Beclin 1、LC3的表达,并用流式细胞术检测低氧与常氧条件下裸鼹鼠成纤维细胞凋亡水平。结果裸鼹鼠皮肤成纤维细胞在低氧与常氧处理24 h时,LC3Ⅱ蛋白的表达量并无明显差异,而低氧处理48 h后LC3Ⅱ蛋白的表达量显著高于常氧组,并且裸鼹鼠成纤维细胞随着低氧培养时间的延长,LC3Ⅱ蛋白表达升高。流式细胞术结果显现低氧处理组裸鼹鼠皮肤成纤维细胞凋亡率显著低于常氧组。结论裸鼹鼠皮肤成纤维细胞可以通过提高自噬水平适应低氧环境,这为今后深入研究裸鼹鼠耐低氧机制提供了参考。

关键词: 裸鼹鼠, 皮肤成纤维细胞, 低氧, 自噬, 凋亡

Abstract: Objective To compare the autophagy and apoptosis level of naked mole rat fibroblasts under normoxia and hypoxia. Methods The naked mole rat fibroblasts were cultured in six-well plates under hypoxic (3% O₂) or normoxia (20% O₂) conditions. The expression of Beclin 1 and LC3 were detected by western blotting and RT-PCR. Annexin V-FITC assay was used to measure the apoptotic cells by flow cytometry. Results The expression levels of LC3Ⅱ were not significantly different between hypoxic and normoxia after 24 hours. An increase was found in the LC3Ⅱ expression when comparing the two groups at 48 hour time points. As the hypoxia time prolonged, the increased expression of LC3 II protein was observed. The results of flow cytometry showed apoptosis rate of hypoxia group was significantly lower than the normal oxygen group. Conclusion Increased autophagy may protect naked mole-rat fibroblasts from hypoxia treatment, which provided insight into the research of hypoxia tolerance mechanism of naked mole rats.

Key words: Heterocephalus glaber, Skin fibroblasts, Hypoxia, Autophagy, Apoptosis

中图分类号:

Q95-33

赵善民, 林丽芳, 肖邦, 王运慧, 宫晨, 张璐, 余琛琳, 汤球, 孙伟, 崔淑芳. 低氧对裸鼹鼠皮肤成纤维细胞自噬水平及凋亡的影响[J]. 实验动物与比较医学, 2014, 34(5): 405-410.

ZHAO Shan-min, LIN Li-fang, XIAO Bang, WANG Yun-hui, GONG Chen, ZHANG Lu, YU Chen-lin, TANG Qiu, SUN Wei, CUI Shu-fang. Comparative Effects of Hypoxia on Autophagy and Apoptosis of Naked Mole Rat Fibroblasts[J]. Laboratory Animal and Comparative Medicine, 2014, 34(5): 405-410.

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低氧对裸鼹鼠皮肤成纤维细胞自噬水平及凋亡的影响

Comparative Effects of Hypoxia on Autophagy and Apoptosis of Naked Mole Rat Fibroblasts

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