基于共聚焦激光显微内镜的海水淹溺肺损伤大鼠肺通透性的实时活体评价  

Real-time assessment of pulmonary permeability in rats with seawater drowing ALI using confocal laser endomicroscopy in vivo

作　　者：任佳博 李春笋[2] 刘金霞 魏元辉 王梓瑞 杨震[2] 陈良安 REN Jiabo;LI Chunsun;LIU Jinxia;WEI Yuanhui;WANG Zirui;YANG Zhen;CHEN Liang'an(Chinese PLA Medical School,Beijing 100853,China;Department of Respiratory and Critical Care Medicine,the First Medical Center,Chinese PLA General Hospital,Beijing 100853,Chin)

机构地区：[1]解放军医学院,北京100853 [2]解放军总医院第一医学中心呼吸与危重症医学科,北京100853

出　　处：《解放军医学院学报》2024年第6期681-688,共8页Academic Journal of Chinese PLA Medical School

基　　金：首都卫生发展科研专项项目(首发2020-1-5011)。

摘　　要：背景急性肺损伤是临床常见急危重症,其核心病理特征是气血屏障的破坏,目前对肺损伤中气血屏障结构及功能改变进行活体评估的研究甚少。目的使用共聚焦激光显微内镜实现大鼠肺的在体可视化成像和海水淹溺肺损伤大鼠肺通透性的实时动态活体评估。方法10只SD大鼠随机分为对照组和海水淹溺(seawater drowning,SW)组,每组5只;SW组通过气管滴注人工海水建立海水淹溺肺损伤模型,造模后3 h进行肺活体观察窗开窗手术,4 h时经股静脉推注0.2 mL异硫氰酸荧光素-葡聚糖(10 mg/mL);对照组不进行海水滴注,其余操作同SW组。使用共聚焦激光显微内镜对右肺下叶进行30 min活体观察,对成像0 min、10 min、20 min、30 min时的共聚焦图像进行染料渗漏指数及荧光分布均匀程度的分析;成像结束后处死大鼠,取肺组织、外周血及肺泡灌洗液,进行组织病理学检查、FITC-Dextran渗漏量检测、肺泡灌洗液蛋白含量检测及肺组织湿/干比测定。结果经静脉给予FITC-Dextran后,共聚焦激光显微内镜下可观察到肺微循环和肺泡结构的实时图像;对照组血管及肺泡结构较清晰,未出现血管外空间荧光强度的明显增加,而SW组血管结构随时间推移逐渐模糊,各时间点SW组的渗漏指数均高于对照组(P<0.05),10 min、20 min、30 min时荧光分布的变异系数均低于对照组(P<0.05);SW组的染料渗漏量、肺泡灌洗液蛋白含量及肺组织湿/干比均高于对照组(P<0.05)。结论海水淹溺造成大鼠肺组织结构破坏,肺通透性增加;利用共聚焦激光显微内镜可实现大鼠肺组织结构的在体可视化成像,通过对共聚焦图像的定量分析,还可实现大鼠气血屏障通透性的实时活体评估。Background Acute lung injury(ALI),characterized by the destruction of the blood–air barrier,is a common clinical emergency.However,in vivo evaluations of the structural and functional changes to the barrier caused by ALI are limited.Objective To explore a real-time visualization method based on confocal laser endomicroscopy(CLE)to examine the permeability of the blood–air barrier in living rats with seawater drowning ALI.Methods Ten SD rats were randomly divided into control group and seawater drowning group(SW group),with 5 rats in each group.The model of seawater drowning ALI was established using the intratracheal instillation of artificial seawater in the SW group.Three hours after modeling,the window-opening surgery was performed;four hours after modeling,FITC-Dextran was injected through a femoral vein catheter.The control group did not carry out seawater instillation,while the rest of the procedure was identical to that of the SW group.The lung’s right inferior lobe was then observed in vivo for 30 minutes using CLE.The leakage index of FITC-Dextran and the uniformity of the fluorescence distribution of the confocal images were then analyzed.Lung tissue,peripheral blood,and bronchoalveolar lavage fluid(BALF)were collected for the histopathological examination and determination of the FITC-Dextran leakage volume,the protein content of BALF,and the wet-to-dry(W/D)weight ratio of the lung tissue,respectively.Results Following the intravenous administration of FITC-Dextran,the real-time images of pulmonary microcirculation and alveolar structure were observed using CLE.In the SW group,the vascular structures grew blurred over time,and there was a significantly higher leakage index than the control group at all time points(P<0.05).Additionally,the variation coefficients of the fluorescence distribution at different time points were lower in the SW group (P<0.05). The leakage volume of FITC-Dextran, the protein content of BALF, and the wet-to-dry weight ratio of the lung tissue were all higher in the SW gr

关 键 词：急性肺损伤 活体成像 共聚焦激光显微内镜 气血屏障 肺通透性 

分 类 号：R563.9[医药卫生—呼吸系统]