摘要
目的:探讨LPS诱导的巨噬细胞来源外泌体对巨噬细胞抗结核分枝杆菌(M.tb)自噬的作用及机制研究。方法:使用THP-1细胞诱导建立H37Ra感染巨噬细胞模型,抽提并鉴定巨噬细胞外泌体;抽提LPS诱导的巨噬细胞外泌体与H37Ra感染巨噬细胞共培养,使用Dil荧光标记外泌体,荧光显微镜观察外泌体吸收情况,采用荧光定量PCR、流式细胞术、Western blotting等方法,明确外泌体对巨噬细胞抗M.tb自噬的影响。结果:成功抽提并鉴定LPS刺激的巨噬细胞外泌体,吸收普通外泌体的巨噬细胞自噬水平无明显变化,吸收LPS诱导的外泌体后的巨噬细胞其自噬水平显著提高,共培养时加入外泌体抑制剂后促进巨噬细胞自噬作用消失;H37Ra感染的巨噬细胞吸收LPS诱导的外泌体后,其自噬水平增强,显著提高对M.tb的杀伤能力及人白细胞介素12的分泌。结论:LPS诱导的巨噬细胞来源外泌体可促进巨噬细胞抗M.tb自噬。
Objective:To investigate the effect and mechanism of LPS-induced macrophage-derived exosomes on macrophage anti-Mycobacterium tuberculosis(M.tb)autophagy.Methods:H37Ra-infected macrophages were induced by THP-1 cells,and macrophage exosomes were extracted and identified;LPS-induced macrophage exosomes were extracted and co-cultured with H37Ra infected macrophages.The exosomes were labeled with Dil fluorescence,and the absorption of exosomes was observed under fluorescence microscope.The effects of exosomes on macrophages′anti-M.tb autophagy were determined by fluorescent quantitative PCR,flow cytometry,Western blotting and other methods.Results:The macrophage exosomes stimulated by LPS were successfully extracted and identified.There was no significant change in the autophagy level of macrophages that absorbed ordinary exosomes.The autophagy level of macrophages that absorbed LPS-induced exosomes was significantly increased.The effect of promoting macrophage autophagy disappeared when exosomes inhibitor was added during coculture;after the macrophages infected with H37Ra absorbed LPS-induced exosomes,the autophagy level was enhanced,and the killing ability of M.tb and the secretion of human interleukin-12 were significantly increased.Conclusions:LPS-induced macrophage-derived exosomes can promote macrophage anti-M.tb autophagy.
作者
方芳
杨珂佳
王瑞祥
钱中清
FANG Fang;YANG Kejia;WANG Ruixiang;QIAN Zhongqing(School of Laboratory Medicine,Bengbu Medical University,Bengbu Anhui 233030,China;Anhui Province Key Laboratory of Immunology in Chronic Diseases,Bengbu Medical University,Bengbu Anhui 233030,China)
出处
《蚌埠医学院学报》
CAS
2024年第4期438-442,447,共6页
Journal of Bengbu Medical College
基金
安徽省高校自然科学研究重点项目(KJ2020A0583)。
关键词
巨噬细胞
外泌体
结核分枝杆菌
自噬
macrophages
exosomes
Mycobacterium tuberculosis
autophagy
