| 深低温暴露诱导的人肾小管上皮细胞自噬体数量特征分析 |
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| 引用本文： | 杨希营,王古岩,陈一萌,张丛雅,杜英杰. 深低温暴露诱导的人肾小管上皮细胞自噬体数量特征分析[J]. 中国分子心脏病学杂志, 2021, 0(1): 3744-3747 |
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| 作者姓名： | 杨希营 王古岩 陈一萌 张丛雅 杜英杰 |
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| 作者单位： | 北京协和医学院国家心血管病中心中国医学科学院阜外医院麻醉科;首都医科大学附属北京同仁医院麻醉科 |
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| 基金项目： | 国家自然科学基金(81770414,81970344)。 |
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| 摘 要： | 目的 通过构建HK2-GFP-LC3稳定细胞系,研究20℃深低温暴露诱导的自噬体数量特征,为明确自噬低温暴露响应和自噬相关药物高通量筛选提供依据.方法 构建稳定表达GFP-LC3融合蛋白的HK2-GFP-LC3细胞系,建立HK2-GFP-LC3细胞体外20℃低温暴露自噬分析模型,用荧光显微镜进行自噬体计数,进行低温暴露...
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| 关 键 词： | 自噬 自噬体 低温暴露 急性肾损伤 |
Quantitative Characteristics of Autophagosomes in Human Renal Tubular Epithelial Cells Induced by Deep Hypothermic Exposure |
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| YANG Xi-ying,WANG Gu-yan,CHEN Yi-meng,ZHANG Cong-ya,DU Ying-jie. Quantitative Characteristics of Autophagosomes in Human Renal Tubular Epithelial Cells Induced by Deep Hypothermic Exposure[J]. Molecular Cardiology of China, 2021, 0(1): 3744-3747 |
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| Authors: | YANG Xi-ying WANG Gu-yan CHEN Yi-meng ZHANG Cong-ya DU Ying-jie |
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| Affiliation: | (Department of Anesthesiology,Fuwai Hospital,National Center for Cardiovascular Disease,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100037,China;Department of Anesthesiology,Beijing Tongren Hospital,Capital Medical University,Beijing 100730,China) |
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| Abstract: | Objective To study the quantitative characteristics of autophagosomes induced by deep hypothermic exposure at 20℃in constructed HK2-GFP-LC3 stable cell lines for the identification of autophagy and the high-throughput screening of autophagy-related agents.Methods HK2-GFP-LC3 cell lines with stable expression of GFP-LC3 fusion protein were constructed to establish cold exposure model at 20Ⅱin vitro for autophagosome analysis.The autophagosomes were counted by fluorescence microscope and the correlation analysis between hypothermic exposure and autophagosomes was conducted subsequently.Meanwhile a linear regression model was established.Results GFP-LC3 spots increased gradually after exposure at 20℃.There was a linear positive correlation between the number of GFP-LC3 spots and the duration of hypothermic exposure,and the fitting formula was Y=1.649×X+17.41.GFP-LC3 spots decreased gradually after rewarming.The trendline of GFP-LC3 spots showed an inverted U-shaped curve in the process of hypothermic exposure and rewarming.Conclusion Hypothermic exposure can activate autophagy in a time-dependent manner in vitro.Rewarming can likely inhibit autophagy. |
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| Keywords: | Autophagy Autophagosome Hypothermic exposure Acute kidney injury |
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