抗凝血酶Ⅲ通过降低肺泡屏障通透性减轻脓毒症急性肺损伤  

AntithrombinⅢreduces sepsis⁃induced acute lung injury by reducing alveolar barrier permeability

作　　者：李东旭 宋振举 王璐[3] 于兵 戴青青 LI Dongxu;SONG Zhenju;WANG Lu;YU Bing;DAI Qingqing(Dept.of Critical Care Medicine,Obstetrics&Gynecology Hospital of Fudan University,Shanghai 200082,China;Dept.of Emergency,Zhongshan Hospital Fudan University&Institute of Emergency Rescue and Critical Care,Fudan University,Shanghai 200032,China;Dept.of Critical Care Medicine,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei,China;Dept.of Cell Biology,Naval Medical University,Shanghai 200433,China)

机构地区：[1]复旦大学附属妇产科医院重症医学科,上海200082 [2]复旦大学附属中山医院急诊科/复旦大学应急救援与急危重症研究所,上海200032 [3]武汉大学人民医院重症医学科,湖北武汉430060 [4]中国人民解放军海军军医大学细胞生物学教研室,上海200433

出　　处：《武汉大学学报(医学版)》2025年第2期153-159,共7页Medical Journal of Wuhan University

基　　金：国家重点研发计划项目(项目号:2021YFC2501800);国家自然科学基金青年科学基金(项目号:82002077);上海市公共卫生体系建设三年行动计划(项目号:GWVI⁃2.1⁃3)。

摘　　要：目的:探讨抗凝血酶Ⅲ(ATⅢ)对脓毒症急性肺损伤肺泡屏障的保护作用。方法:将雄性C57BL/6J小鼠随机分成3组,①脓毒症组(LPS组):将小鼠饥饿16 h后,腹腔注射20 mg/kg脂多糖(LPS);②脓毒症+ATⅢ组(LPS+ATⅢ组):LPS注射2 h后,尾静脉注射750 IU/kg ATⅢ;③对照组(Control组):参照LPS和ATⅢ的给药途径与时间分别注射等体积生理盐水。在ATⅢ给药24 h后收集血清、肺泡灌洗液以及肺组织等标本,采用HE染色结合肺损伤评分评价肺组织的形态结构与损伤程度;采用肺湿/干重比(W/D)、肺泡灌洗液(BALF)中的细胞数量与蛋白量以及FITC-Dextran渗透实验测量肺泡屏障通透性;采用免疫组织学染色检测肺组织细胞间紧密连接组成蛋白的表达量。结果:LPS组的肺损伤评分、肺湿/干重比(W/D值)、肺泡屏障通透性均较Control组显著升高(P<0.01),细胞间紧密连接的组成蛋白ZO-1、Occludin和Claudin3的表达水平均显著降低(P<0.001)。而LPS+ATⅢ组的肺损伤评分、W/D值、肺泡屏障通透性较LPS组均降低(P<0.05),ZO-1、Occludin和Claudin3表达水平升高(P<0.01)。结论:ATⅢ增加细胞间紧密连接组成蛋白的表达,进而降低肺泡屏障的通透性,从而减轻脓毒症急性肺损伤。Objective:To explore the protective effect of antithrombinⅢ(ATⅢ)on the alveolar barrier in acute lung injury induced by sepsis.Methods:Male C57BL/6J mice were randomly divided into three groups,①sepsis group(LPS group):after fasting the mice for 16 hours,20 mg/kg lipopolysaccharide(LPS)was injected intraperitoneally;②sepsis treated with ATⅢgroup(LPS+ATⅢgroup):750 IU/kg ATⅢwas injected via the tail vein 2 hours after LPS injection;③control group:equal volume of saline were injected according to the strategies of the administration of LPS and ATⅢ.The serum,bronchoalveolar lavage fluid(BALF),and lung tissue samples were collected 24 hours after ATⅢad-ministration.HE staining combined with lung injury scoring was used to assess the morphological structure and degree of damage in lung tissues;the lung wet/dry weight ratio(W/D),cell count and protein content in BALF,and FITC-Dextran permeability assay were employed to measure pulmo-nary barrier permeability;immunohistochemical staining was utilized to detect the expression levels of tight junction proteins in lung tissues.Results:The lung injury score,lung W/D value and the perme-ability of the alveolar barrier in the LPS group were significantly increased as compared with those in the control group(P<0.01);the expression levels of the components of tight junctions,ZO-1,occludin,and Claudin3 were significantly reduced(P<0.001).However,the lung injury score,W/D value,and the permeability of the alveolar barrier in the LPS+ATⅢgroup were lower than those in the LPS group(P<0.05),and the expression level of ZO-1,occludin,and Claudin3 was higher(P<0.01).Conclusion:ATⅢreduces the permeability of the alveolar barrier by increasing the expression of tight junction proteins,thus alleviating acute lung injury induced by sepsis.

关 键 词：脓毒症 急性肺损伤 抗凝血酶Ⅲ 肺泡屏障 

分 类 号：R56[医药卫生—呼吸系统]