摘要目的:探讨自噬对小鼠巨噬细胞吞噬创伤弧菌能力的影响。方法:采用Western blot检测创伤弧菌感染RAW264.7细胞和BMMφ细胞自噬标志物的表达变化；构建带荧光蛋白标记的创伤弧菌(Vv-GFP)感染RAW264.7细胞和BMMφ细胞，共聚焦显微镜观察巨噬细胞吞噬后的创伤弧菌与LC3蛋白共定位情况；采用Bafilomycin A1抑制自噬，Western blot检测自噬标志物的表达变化，流式细胞术检测巨噬细胞吞噬清除创伤弧菌能力的改变。结果:创伤弧菌感染诱导RAW264.7细胞和BMMφ细胞Atg7、Atg12、Atg16L1和LC3Ⅱ的表达显著上调，p62蛋白表达显著下调；巨噬细胞吞噬的创伤弧菌与自噬体形成膜标志物LC3B发生共定位；创伤弧菌感染促进了自噬体形成和自噬底物降解，促进巨噬细胞对创伤弧菌的吞噬作用，而自噬抑制剂Bafilomycin A1则通过抑制自噬/溶酶体降解过程，导致Atg7、Atg12、Atg16L1和LC3Ⅱ表达累积，p62表达上调，阻断自噬流，进而减弱巨噬细胞对创伤弧菌吞噬清除的能力。结论:创伤弧菌感染诱导巨噬细胞发生自噬，有利于增强巨噬细胞吞噬清除创伤弧菌的能力。

abstractsObjective:To investigate the role of autophagy in the regulatory of phagocytosis in Vibrio vulnificus ( V. vulnificus)-infected murine macrophages. Methods:The expression of cellular autophagy-related proteins in PBS-treated and V. vulnificus-infected RAW264.7 and BMMφ cells was detected by Western blot. The co-localization of V. vulnificus-GFP and LC3Ⅱ protein in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells were detected using confocal microscopy. The phagocytosis of V. vulnificus in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells with or without autophagy inhibition using Bafilomycin A1 was detected by flow cytometry. Results:The up-regulated levels of Atg7, Atg12 and Atg16L1 proteins, increased LC3Ⅱ/actin ratio, as well as down-regulated p62 protein levels were significantly detected in V. vulnificus-infected RAW264.7 and BMMφ cells. The co-localization of V. vulnificus-GFP and LC3Ⅱ protein was clearly observed in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells. Enhanced phagocytosis of V. vulnificus and increased autophagy were exhibited in V. vulnificus-GFP-infected RAW264.7 and BMMφ cells, while weakened phagocytosis, accumulation of Atg7, Atg12, Atg16L1, LC3Ⅱ and p62 protein levels, as well as blocking autophagy flux were detected in those cells within autophagy inhibition using Bafilomycin A1. Conclusion:Autophagy induced by V. vulnificus infection could promote phagocytosis of V. vulnificus in macrophages.