现代生物医学进展biomed.cnjournals.comProgressinModernBiomedicineVol.23NO.2JAN.2023doi:10.13241/j.cnki.pmb.2023.02.002雷公藤甲素诱导肝细胞选择性自噬的形态学观察*蒋杨雨江始源曹丹王海燕邹昊靖李健△（北京中医药大学中医学院组胚教研室北京102488）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，Westernblot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。关键词：选择性自噬；雷公藤甲素；肝细胞；形态学中图分类号：R-33；Q593.1文献标识码：A文章编号：1673-6273（2023）02-206-05MorphologicObservationofHepatocyteSelectivelyAutophagyInducedbyTriptolide*JIANGYang-yu,JIANGShi-yuan,CAODan,WANGHai-yan,ZOUHao-jing,LIJian△(DepartmentofHistologyandEmbryology,SchoolofTraditionalChineseMedicine,BUCM,Beijing,102488,China)ABSTRACTObjective:Toobservethelevelofhepatocyteselectiveautophagyinducedbytriptolide.Methods:Anautophagytrac-ingmodelinhepatocyteswasestablishedbyrapidtailveininjectionofGFP-LC3plasmidsintomicetreatedwithtriptolidetoobservethehepatocyteautophagylevelinducedbytriptolideinvivo.GFP-LC3stableexpressedL02cellsweretransfectedwithRFP-P62plasmid,andtheselectiveautophagyprofileofL02andLC3-P62complexinducedbytriptolideweredynamicallyobservedbylivecellimagingsystemandfluorescencemicroscopy.Results:Theresultsofinvivoexperimentsshowedthattriptolidesignificantlyinducedautophagyinhepatocytes.AndasimilartrendontheproteinexpressionofLC3IIandP62wasdemonstratedbyWesternblot.Theresultsoflivecellimagingsystemandfluorescencemicroscopyshowedtheco-localizationofLC3andP62suggesti...
