本研究探讨了源自 pH 7.4 和 9.0.E 培养基的粪肠球菌膜囊泡 (MV) 的促炎作用。通过离心获得粪肠球菌MV并通过尺寸排阻色谱法纯化。对粪肠球菌 MV 进行蛋白质组分析以研究其成分。将 THP-1 巨噬细胞暴露于粪肠球菌 MV，并使用 ELISA 和免疫印迹评估炎症细胞因子和蛋白质。采用ELISA法检测腹腔注射粪肠球菌MV的小鼠血清中的炎性细胞因子，并采用流式细胞术研究脾细胞的免疫表型。蛋白质组学分析显示中性和碱性条件下获得的粪肠球菌MV中有196个蛋白，110个蛋白。碱性 pH 值导致蛋白质上调，79 种蛋白质下调。粪肠球菌MV以浓度依赖性方式诱导炎症因子白细胞介素（IL）-1β、IL-6和肿瘤坏死因子-α的分泌。免疫印迹显示，粪肠球菌 MV 增加了 pro-IL-1β、核因子-κBp65 和 Toll 样受体 2 的表达。体内研究表明，粪肠球菌 MV 显着促进小鼠血清中 IL-1β 的分泌，同时炎症脾脏中的细胞被激活。在 pH 9.0 下获得的粪肠球菌 MV 比在中性 pH 下获得的 MV 显示出更强的促炎作用。粪肠球菌产生的MV携带与毒力因子相关的特定蛋白质，这些MV可以在体外和体内促进炎症。碱性条件下获得的粪肠球菌MV具有更强的促炎作用。版权所有©2024。由Elsevier Inc.出版。

The present study explored the proinflammatory impact of Enterococcus faecalis membrane vesicles (MVs) derived from culture medium at pH 7.4 and 9.0.E. faecalis MVs were obtained by centrifugation and purified by size exclusion chromatography. Proteomic analyses were carried out on E. faecalis MVs to investigate their components. THP-1 macrophages were exposed to E. faecalis MVs, and the inflammatory cytokines and proteins were evaluated using ELISA and immunoblotting. The inflammatory cytokines in the serum of mice with intraperitoneal injection of E. faecalis MVs were evaluated by ELISA, and immunophenotyping of spleen cells was investigated with flow cytometry.Proteomic analysis revealed 196 proteins in E. faecalis MVs obtained under neutral and alkali conditions, 110 proteins were upregulated and 79 proteins were downregulated by alkaline pH. E. faecalis MVs induced secretion of inflammatory factors interleukin (IL)-1β, IL-6 and tumor necrosis factor-α in a concentration-dependent manner. Immunoblotting revealed that E. faecalis MVs increased expression of pro-IL-1β, nuclear factor-κBp65, and Toll-like receptor 2. In vivo studies demonstrated that E. faecalis MVs significantly promoted secretion of IL-1β in mouse serum, while inflammatory cells were activated in the spleen. E. faecalis MVs obtained at pH 9.0 showed stronger proinflammatory effects than those obtained under neutral pH.E. faecalis produce MVs that carry specific proteins associated with virulence factors, and these MVs can promote inflammation in vitro and in vivo. E. faecalis MVs obtained under alkaline conditions have a stronger proinflammatory effect.Copyright © 2024. Published by Elsevier Inc.