摘要
目的:本研究旨在探讨干扰素基因刺激因子(STING)对小鼠急性肺损伤修复过程的作用,并深入研究其对小鼠巨噬细胞胞葬功能的影响。方法:选取25只10~12周龄的C57BL/6小鼠,并随机分为D0组、D1组、D3组、D5组、D7组,每组5只,分别给予气管内注射0.5 mg/kg脂多糖(LPS)0、1、3、5、7 d后收集小鼠样本。另外,分别选择野生型STING^(+/+)和STING基因缺失型(STING^(-/-))小鼠各5只,分为STING^(+/+)+LPS组和STING^(-/-)+LPS组,同样给予气管内注射0.5 mg/kg LPS诱导5 d后收取小鼠样本。通过苏木素-伊红(HE)染色观察小鼠肺组织的损伤病理变化,并检测小鼠肺泡灌洗液细胞数、中性粒细胞数、蛋白浓度来评估肺损伤程度;经小鼠气管注射凋亡的中性粒细胞,并于3 h后收集肺泡灌洗液细胞进行流式细胞术检测,观察巨噬细胞的胞葬功能;采用免疫荧光染色法对肺组织进行TUNEL和F4/80双染评估胞葬作用;并采用巴氏染色法观察肺泡巨噬细胞的胞葬现象。结果:C57BL/6小鼠诱导急性肺损伤后3 d达到肺损伤的顶峰,5 d后开始修复,并在不治疗的情况下,于7 d内修复基本完成。小鼠急性肺损伤5 d后,与STING^(+/+)+LPS组小鼠相比,STING^(-/-)+LPS组小鼠的肺损伤评分、肺泡灌洗液总蛋白浓度、肺泡灌洗液总细胞数、肺泡灌洗液中性粒细胞数均较STING^(+/+)+LPS组更低(t=3.257、2.926、3.946、2.669,P=0.012、0.019、0.004、0.028)。流式细胞术及免疫荧光染色检测结果显示,STING^(-/-)+LPS组小鼠肺泡巨噬细胞胞葬率均显著高于STING^(+/+)+LPS组(t=3.143、6.963,P=0.016、<0.001)。结论:STING缺失可能通过促进巨噬细胞胞葬功能,从而调控急性肺损伤的修复过程。本研究有望为深入理解STING在肺部损伤和修复中的作用提供新的见解。
Objective:To investigate the impact of the stimulator of interferon genes(STING)on the repair process of acute lung injury in mice and its influence on macrophages efferocytosis.Methods:Twenty-five C57BL/6 mice aged 10-12 weeks were randomly divided into a D0 group,a D1 group,a D3 group,a D5 group and a D7 group,with five mice in each group,and the mice samples were collected after 0,1,3,5 and 7 days of an intratracheal injection of 0.5 mg/kg lipopolysaccharide(LPS)respectively.In addition,five STING wild-type(STING^(+/+))mice and five STING knockout(STING^(-/-))mice were selected and divided into a STING^(+/+)+LPS group and a STING^(-/-)+LPS group,and the mice samples were collected after 5 days of an intratracheal injection of 0.5 mg/kg LPS.Hematoxylin eosin(HE)staining was employed to observe the pathological changes in lung tissue,and the cell count,neutrophil count and protein concentration in alveolar lavage fluid were detected to access the degree of lung injury.Apoptotic neutrophils were injected into the trachea of mice,and alveolar lavage fluid cells were collected 3 hours later for flow cytometry to determine the efferocytosis rate of macrophages.TUNEL and F4/80 double staining of lung tissues were conducted via immunofluorescence to evaluate efferocytosis.The phenomenon of macrophage efferocytosis was observed through Pap staining.Results:The C57BL/6 mice reached the peak of lung injury 3 days after induction of acute lung injury and began to repair after 5 days,and in the absence of treatment,the repair was basically completed within 7 days.After 5 days of acute lung injury,the lung tissue injury score and the protein concentration,total cell count and neutrophil count in alveolar lavage fluid of mice in the STING^(-/-)+LPS group were lower than those in the STING^(+/+)+LPS group(t=3.257,2.926,3.946,2.669;P=0.012,0.019,0.004,0.028).Flow cytometry and immunofluorescence staining showed that the efferocytosis rates of alveolar macrophages in the STING^(-/-)+LPS group were significantly higher than thos
作者
李璐璐
马利红
金佳佳
谷伟
Li Lulu;Ma Lihong;Jin Jigjia;Gu Wei(Departmentof Respiration,NanjingFirst Hospital,Nanjing Medical University,Nanjing 210006,China;Department of Respiratory and Critical Care Medicine,Wuxi Second People's Hospital,Wuxi 214086,China;Department of Respiratory and Critical Care Medicine,Jinling Hospital,Nanjing Medical University,Nanjing 210002,China)
出处
《中华危重症医学杂志（电子版）》
CAS
CSCD
2024年第2期97-103,共7页
Chinese Journal of Critical Care Medicine:Electronic Edition
基金
国家自然科学基金项目(82100095)。
