| 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].现代生物医学进展英文版,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 |
| Received:April 23, 2022 Revised:May 18, 2022 |
| 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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