【摘要】 目的:鉴定海分枝杆菌感染巨噬细胞过程中分泌的细菌效应蛋白，为揭示其胞内生存机制及解决非结核分枝杆菌耐药问题提供理论基础。方法:采用海分枝杆菌感染RAW 264.7小鼠巨噬细胞，通过液相色谱-串联质谱技术系统鉴定细菌来源的分泌蛋白，并结合生物信息学方法进行基因本体论(gene ontology,GO)富集分析、京都基因与基因百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路分析、蛋白质相互作用(protein-protein interaction,PPI)分析。结果:从海分枝杆菌感染的RAW 264.7细胞上清液中鉴定到1629种细菌分泌蛋白。GO主要富集到细菌的代谢过程与应激反应等生物过程，以及酶活等分子功能。KEGG通路富集分析将RNA降解通路定位为最显著的富集通路，该通路涉及9种分泌蛋白。PPI分析筛选的枢纽分子主要与核糖体功能有关。结论:本研究首次系统鉴定了海分枝杆菌在宿主巨噬细胞内细菌分泌蛋白的种类及其功能特征，这些蛋白不仅显著富集于细菌代谢及RNA降解等关键通路。同时，海分枝杆菌的MMAR＿1032、MMAR＿1052等效应蛋白作为核心枢纽分子可能在非结核分枝杆菌感染的发生与发展中扮演重要角色。更多还原

【Abstract】 Objective: Dentifying bacterial effector proteins secreted by Mycobacterium marinum during infection of macrophages provides a theoretical foundation for elucidating its intracellular survival mechanisms and addressing drug resistance issues in non-tuberculous mycobacteria. Methods: Mycobacterium marinum infection of RAW 264.7 mouse macrophages was used to identify bacterial-derived secreted proteins systematically via liquid chromatography-tandem mass spectrometry. This was complemented by bioinformatics approaches,including gene ontology enrichment analysis,Kyoto Encyclopedia of Genes and Genomes pathway analysis,and protein-protein interaction analysis. Results: A total of 1,629 bacterial secreted proteins were identified in the supernatant of RAW 264.7 cells infected with Mycobacterium marinum. GO terms predominantly enrich biological processes such as bacterial metabolism and stress responses,as well as molecular functions including enzyme activity. KEGG pathway enrichment analysis identified the RNA degradation pathway as the most significantly enriched pathway,involving nine secreted proteins. The key molecules identified through PPI analysis primarily relate to ribosomal function. Conclusion: This study systematically elucidates for the first time the types of proteins secreted by Mycobacterium marinum within host macrophages and their functional characteristics. These proteins are significantly enriched in key pathways such as bacterial metabolism and RNA degradation. Concurrently,effector proteins of Mycobacterium marinum,such as MMAR＿1032 and MMAR＿1052,may function as pivotal hub molecules in the pathogenesis and progression of non-tuberculous mycobacterial infections.更多还原