水通道蛋白1作为急性呼吸窘迫综合征肺部炎症水平的新生物标志物探索研究  

An exploratory research of aquaporin-1 as a new biomarker of pulmonary inflammation in acute respiratory distress syndrome

作　　者：郭星星 张欢欢 李静 蔡志贵 剡旺 宋立强 GUO Xingxing;ZHANG Huanhuan;LI Jing;CAI Zhigui;YAN Wang;SONG Liqiang(Department of Pulmonary and Critical Care Medicine,Xijing Hospital,Air Force Medical University,Xi'an 710032,China)

机构地区：[1]空军军医大学西京医院呼吸与危重症医学科,陕西西安710032

出　　处：《空军军医大学学报》2024年第4期418-424,共7页Journal of Air Force Medical University

基　　金：陕西省自然科学基础研究重点项目(2022JZ-58);空军军医大学军事医学提升计划项目(2021JSTS36);空军军医大学西京医院学科助推计划项目(XJZT21CZ06)。

摘　　要：目的探讨急性呼吸窘迫综合征(ARDS)肺组织水通道蛋白1(AQP1)表达水平的时效规律以及与肺损伤程度、经典炎症指标的相关性,旨在为ARDS寻找新的生物标志物及干预靶点。方法脂多糖(LPS)滴鼻制备ARDS小鼠模型,每日称量小鼠体质量,取6 h、1 d、3 d和7 d四个时间点肺组织和肺泡灌洗液,HE分析肺组织病理变化,ELISA和qPCR检测炎症因子IL-1β、IL-6和TNF-α的表达水平,流式细胞术分析中性粒细胞、肺泡巨噬细胞、嗜酸性粒细胞、单核细胞等天然免疫细胞数量及比例,qPCR和Western blotting检测AQP1 mRNA和蛋白表达水平,Pearson法分析AQP1与炎性细胞相关性。结果LPS刺激6 h后小鼠肺组织有炎性细胞浸润,至第3日出现严重肺损伤,第7日有所恢复;炎症因子IL-1β、IL-6和TNF-αmRNA和蛋白水平持续升高至第3日,第7日降低(P<0.05);中性粒细胞比例显著升高,第7日降低,肺泡巨噬细胞和嗜酸性粒细胞、单核细胞比例持续降低至第3日,第7日升高(P<0.05);AQP1 mRNA和蛋白表达水平持续降低(P<0.05);AQP1表达水平与四种炎性细胞显著相关(P<0.01)。结论ARDS肺组织AQP1表达水平与肺损伤程度及炎症指标呈显著负相关,可作为肺损伤的新生物标志物,进一步揭示AQP1可能扮演的保护性角色具有潜在临床价值。Objective To investigate the aging rule of aquaporin-1(AQP1)expression level in lung tissue in acute respiratory distress syndrome(ARDS),and its correlation with the degree of lung injury and classic inflammatory indicators,so as to find novel biomarkers and intervention targets for ARDS.Methods A mouse model of ARDS was prepared by intranasal instillation of lipopolysaccharide(LPS).The body mass of the mice was recorded daily,and lung tissue and bronchoalveolar lavage fluid were collected at 6 h,1 d,3 d,and 7 d.HE was used to analyze the pathological changes in lung tissue.ELISA and qPCR were used to detect the levels of inflammatory factors IL-1β,IL-6,and TNF-α.Flow cytometry was used to analyze the number and proportion of innate immune cells,such as neutrophils,alveolar macrophages,eosinophils,and monocytes.qPCR and Western blotting were used to detect AQP1 mRNA and protein expression levels.Pearson's method was used to analyze the correlation between AQP1 and inflammatory cells.Results After LPS stimulation for 6 h,there was inflammatory cell infiltration in the lung tissue of mice,severe lung injury appeared on day 3,and recovered on day 7.The mRNA and protein levels of inflammatory factors IL-1β,IL-6,and TNF-αincreased until day 3 and decreased on day 7(P<0.05).The proportion of neutrophils was significantly increased and decreased on day 7,while the proportion of alveolar macrophages,eosinophils,and monocytes continued to decrease until day 3 and increased on day 7(P<0.05).AQP1 mRNA and protein expression levels continuously decreased(P<0.05).The expression level of AQP1 was significantly correlated with four kinds of inflammatory cells(P<0.01).Conclusion The expression level of AQP1 in lung tissue in ARDS is significantly negatively correlated with the degree of lung injury and inflammatory indicators,which can be used as a new biomarker of lung injury,and further reveal the possible protective role of AQP1 with potential clinical value.

关 键 词：急性呼吸窘迫综合征 水通道蛋白1 脂多糖 炎性细胞 炎性因子 

分 类 号：R563[医药卫生—呼吸系统]