蒋杨雨,江始源,曹 丹,王海燕,邹昊靖,李 健.雷公藤甲素诱导肝细胞选择性自噬的形态学观察[J].,2023,(2):206-210 |
雷公藤甲素诱导肝细胞选择性自噬的形态学观察 |
Morphologic Observation of Hepatocyte Selectively Autophagy Induced by Triptolide |
投稿时间:2022-04-23 修订日期:2022-05-18 |
DOI:10.13241/j.cnki.pmb.2023.02.002 |
中文关键词: 选择性自噬 雷公藤甲素 肝细胞 形态学 |
英文关键词: Selectively autophagy Triptolide Hepatocyte Morphology |
基金项目:国家自然科学基金面上项目(81873099) |
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中文摘要: |
摘要 目的:观察雷公藤甲素诱导肝细胞选择性自噬的水平。方法:在雷公藤甲素给药处理小鼠的尾静脉高压注射GFP-LC3质粒,制备肝细胞自噬示踪模型,观察雷公藤甲素在动物体内诱导肝细胞自噬的水平。在GFP-LC3稳定表达的L02细胞株中转入RFP-P62质粒,用活细胞工作站和荧光显微镜动态观察雷公藤甲素诱导L02细胞选择性自噬的轮廓,同时也观察细胞自噬复合体LC3-P62的变化。结果:动物实验结果表明,雷公藤甲素可显著诱导肝细胞自噬体的形成,Western blot结果显示P62蛋白和LC3II蛋白的表达趋势一致。活细胞动态观察及免疫荧光双标记实验结果表明LC3-P62存在共定位,提示雷公藤甲素可诱导肝细胞自噬流的形成。结论:雷公藤甲素可诱导肝细胞选择自噬,其生物学意义可能与肝细胞损伤后修复相关。 |
英文摘要: |
ABSTRACT Objective: To observe the level of hepatocyte selective autophagy induced by triptolide. Methods: An autophagy tracing model in hepatocytes was established by rapid tail vein injection of GFP-LC3 plasmids into mice treated with triptolide to observe the hepatocyte autophagy level induced by triptolide in vivo. GFP-LC3 stable expressed L02 cells were transfected with RFP-P62 plasmid, and the selective autophagy profile of L02 and LC3-P62 complex induced by triptolide were dynamically observed by live cell imaging system and fluorescence microscopy. Results: The results of in vivo experiments showed that triptolide significantly induced autophagy in hepatocytes. And a similar trend on the protein expression of LC3II and P62 was demonstrated by Western blot. The results of live cell imaging system and fluorescence microscopy showed the co-localization of LC3 and P62 suggesting that the triptolide could induce autophagy flux. Conclusion: Triptolide induced selective autophagy in hepatocytes and its biological function might be associated with hepatocyte damage repairing after injury. |
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