肺泡巨噬细胞来源的微囊泡对肺损伤的作用  

The effect of alveolar microvesicles derived from macrophages on lung injury

作　　者：张蓝予 秦春妮[1] 梁影 陈书弘 黑飞龙[1] Zhang Lanyu;Qin Chunni;Liang Ying;Chen Shuhong;Hei Feilong(Department of Cardiopulmonary Bypass,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,100037 Beijing,China)

机构地区：[1]中国医学科学院,北京协和医学院国家心血管病中心,心血管疾病国家重点实验,室阜外医院体外循环中心,北京100037

出　　处：《中国体外循环杂志》2021年第2期108-111,共4页Chinese Journal of Extracorporeal Circulation

基　　金：国家自然科学基金(81670077);北京协和医学院创新基金(2019-1002-50)。

摘　　要：目的探究肺泡巨噬细胞来源微囊泡(microvesicles,MVs)对肺损伤的作用。方法培养小鼠肺泡巨噬细胞,通过梯度离心法获取磷酸缓冲盐溶液(phosphate buffer saline,PBS)或脂多糖刺激的肺泡巨噬细胞来源的MVs,并经透射电镜、纳米粒径追踪技术及蛋白免疫印迹进行鉴定。经气管内给予肺泡巨噬细胞来源的MVs,观察小鼠肺组织变化情况,并经蛋白免疫印迹探究肺组织中紧密连接相关蛋白的表达变化。结果透射电镜发现MVs呈现sharp结构,纳米粒径追踪显示MVs粒径集中于100~200 nm,并且其表达CD63、CD68及ALIX标志物。苏木精-伊红染色发现脂多糖刺激的肺泡巨噬细胞来源的MVs可刺激小鼠发生肺损伤,并降低紧密连接相关蛋白闭合小环蛋白,咬合蛋白及闭合蛋白的表达。结论炎性肺泡巨噬细胞来源的MVs可导致肺损伤并降低肺部紧密连接相关蛋白的表达,肺泡巨噬细胞来源的MVs参与肺损伤的发生发展。Objective To explore the effect of microvesicles(MVs)derived from alveolar macrophages on lung injury.Methods Mouse alveolar macrophages were cultured.MVs derived from alveolar macrophages stimulated by PBS or lipopolysaccharide were obtained by gradient centrifugation and identified by transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA)and western blotting.MVs derived from alveolar macrophages were administered intratracheally to observe the changes in mouse lung tissue.The expressions of tight junction-related proteins in lung tissue were explored by western blotting.Results TEM showed that the MVs exhibited a sharp structure,and NTA showed that the MVs were concentrated in 100-200nm.The CD63,CD68 and ALIX markers were expressed on MVs.Hematoxylin-eosin staining showed that MVs derived from lipopolysaccharide-stimulated alveolar macrophages could induce lung injury in mice and reduced the expressions of tight junction-related proteins ZO-1,Occludin and Claudin-5.Conclusion MVs derived from inflammatory alveolar macrophages could induce lung injury and reduce the expressions of tight junction-related proteins in lungs.MVs derived from alveolar macrophages participate in the development of lung injury.

关 键 词：肺泡巨噬细胞 肺损伤 微囊泡 紧密连接相关蛋白 

分 类 号：R563[医药卫生—呼吸系统]