Sestrin2对热暴露肺上皮细胞凋亡的干预作用及其机制*

doi:10.12047/j.cjap.5741.2019.060

中国应用生理学杂志 ›› 2019, Vol. 35 ›› Issue (4): 289-292.doi: 10.12047/j.cjap.5741.2019.060

• 研究论文 • 下一篇

Sestrin2对热暴露肺上皮细胞凋亡的干预作用及其机制^*

杲修杰⁺, 王尚⁺, 刘伟丽, 王坤, 陈照立^△, 王新兴^△

军事科学院军事医学研究院环境医学与作业医学研究所, 天津 300050

收稿日期:2018-08-27 出版日期:2019-07-28 发布日期:2019-11-06
通讯作者: ^△,Tel: 02284655206; E-mail: wxxemail@sina.cn, zhaolichen@126.com.
基金资助:
^*国家自然科学基金面上项目(81372124);部队重点项目(18CXZ044, BWS16J010, AWS16J004)

The effects of Sestrin2 on apoptosis of heat-exposed lung epithelial cells and its mechanism

GAO Xiu-jie⁺, WANG Shang⁺, LIU Wei-li, WANG Kun, CHEN Zhao-li^△, WANG Xin-xing^△

Environmental Medicine Laboratory, Institute of Environmental and Operational Medicine, Academy of Military Medical Sciences, Tianjin 300050, China

Received:2018-08-27 Online:2019-07-28 Published:2019-11-06

摘要/Abstract

摘要： 目的:探讨Sestrin2蛋白对热暴露肺上皮细胞凋亡的干预作用及其作用机制。方法:体外培养的Beas-2B细胞分为对照组(37℃)和热暴露组(39℃、40℃和41℃),在上述温度中暴露不同时间(0、3、6和12 h),胰酶消化后收集细胞,分别通过Western blot、荧光分光光度计、流式细胞仪等方法检测细胞中的Sestrin2、超氧化物歧化酶(SOD)、活性氧自由基(ROS)表达水平,细胞线粒体膜电位及细胞凋亡率。基因序列克隆入高表达质粒pcDNA 3.1⁺中,采用Lipfectamine 2000方法转染Beas-2B细胞,构建Sestrin2和SOD高表达细胞,观察细胞线粒体膜电位及细胞凋亡等指标的变化。结果:随着暴露温度的升高,与对照组相比,热暴露组细胞Sestrin2蛋白表达水平下降。在41℃热暴露Beas-2B细胞,不同时间点ROS水平显著上升,线粒体膜电位显著下降,细胞凋亡率增加。Sestrin2和SOD高表达细胞,在41℃暴露条件下,与对照组比较,ROS表达水平显著降低,线粒体膜电位下降幅度减小,热暴露导致细胞凋亡率降低。结论: Sestrin2能够通过线粒体膜电位和SOD缓解热暴露引起肺上皮细胞的凋亡,对Beas-2B细胞具有保护作用。

关键词: Sestrin2, 热暴露, 氧化应激, 细胞凋亡, Beas-2B细胞

Abstract: Objective: To investigate the protective effects of Sestrin2 protein on lung epithelial Beas-2B cells in the heat-exposure environment and its mechanism. Methods: Lung epithelial Beas-2B cells were cultured at 37℃, 39℃, 40℃ and 41℃ respectively. Cells were harvested at different times (0, 3, 6 and 12 h) after pancreatin digestion. The expressions of Sestrin2, superoxide dismutase(SOD), reactive oxygen species(ROS), cell mitochondrial membrane potential and apoptosis rate of cells were detected by Western blot, fluorescence spectrophotometer and flow cytometry, respectively. Gene expression sequence was cloned into high expression plasmid pcDNA3.1⁺. Beas-2B cells were transfected by Lipfectamine 2000 to construct Sestrin2 and SOD high expression cells. The changes of mitochondrial membrane potential and cell apoptosis were observed in the Sestrin2 and SOD high expression cells. Results: With the increase of temperature, the expression level of Sestrin2 protein in heat treatment group was decreased compared with the control group. When Beas-2B cells were exposed to 41℃, the ROS level was increased, mitochondrial membrane potential was decreased significantly and apoptosis rate was increased at different time points. After high expression of Sestrin2 and SOD in the Beas-2B cells, the expression level of ROS was decreased and the change tendency of mitochondrial membrane potential was decreased, and the apoptosis rate was reduced at 41℃ exposure. Conclusion: Sestrin2 can alleviate the apoptosis of lung epithelial cells induced by heat exposure through mitochondrial membrane potential and SOD, which has protective effect on lung epithelial Beas-2B cells.

Key words: Sestrin2, heat exposure, oxidative stress, apoptosis

杲修杰, 王尚, 刘伟丽, 王坤, 陈照立, 王新兴. Sestrin2对热暴露肺上皮细胞凋亡的干预作用及其机制^*[J]. 中国应用生理学杂志, 2019, 35(4): 289-292.

GAO Xiu-jie, WANG Shang, LIU Wei-li, WANG Kun, CHEN Zhao-li, WANG Xin-xing. The effects of Sestrin2 on apoptosis of heat-exposed lung epithelial cells and its mechanism[J]. CJAP, 2019, 35(4): 289-292.

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Sestrin2对热暴露肺上皮细胞凋亡的干预作用及其机制^*

The effects of Sestrin2 on apoptosis of heat-exposed lung epithelial cells and its mechanism

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