小鼠肺爆震伤蛋白质组学研究  

Study on proteomic changes of lung blast injury in mice

作　　者：解晓荟 佟昌慈 丛培芳 刘颖 柳云恩 金红旭 Xie Xiaohui;Tong Changci;Cong Peifang;Liu Ying;Liu Yun'en;Jin Hongxu(Emergency Medicine Department,General Hospital of Northern Theater Command,Laboratory of Rescue Center of Severe Wound and Trauma PLA,General Hospital of Shenyang Military Command,Shenyang 110016,China)

机构地区：[1]北部战区总医院急诊医学部,全军重症(战)创伤救治中心实验室辽宁省重症创伤和器官保护重点实验室,沈阳110116

出　　处：《中华急诊医学杂志》2021年第5期520-525,共6页Chinese Journal of Emergency Medicine

基　　金：辽宁省自然科学基金指导计划(201602771,20170540947);辽宁省重点研发资助计划(2019010205)。

摘　　要：目的通过检测不同时间点肺爆震伤小鼠肺蛋白质组学变化,探究肺爆震伤损伤机制。方法60只健康雄性C57BL/6小鼠,随机(随机数字法)分为对照组、爆震后12 h组、24 h组、48 h组、72 h组及1周组(每组10只)。在北部战区总医院动物实验室进行实验,利用自主研发的精准爆震装置建立小鼠肺爆震伤模型;通过大体观察和HE染色评价肺组织损伤情况;基于液相色谱-串联质谱联用(liquid chromatography-tandem mass spectrometry,LC-MS/MS)的蛋白质组学技术对小鼠肺组织蛋白进行定量分析,筛选出差异表达蛋白,在此基础上利用生物信息学工具分析蛋白质组学变化。结果肺爆震伤后,小鼠肺组织表面可观察到散在出血点,且随时间延长逐渐增多,24 h出血最为严重。HE染色可见爆震后12 h和24 h的肺泡正常组织结构消失,肺泡腔内弥漫性出血,大量炎细胞浸润,肺间质渗出液增多,肺泡壁增厚,肺泡腔萎陷融合。LC-MS/MS共鉴定到6861个蛋白质,定量到608个差异表达蛋白,其中在12、24、48、72 h及1周分别有227个、140个、202个、258个和71个差异蛋白。基因本体论(gene ontology,GO)分析得到包括细胞粘附、细胞外基质组织及胶原原纤维组织等130个生物进程子类,外泌体、细胞外基质及细胞质等66个细胞组分子类,以及细胞外基质结构组成、肌动蛋白结合及抗氧化活性等43个分子功能子类。KEGG分析共得到细胞外基质受体相互作用、黏着斑及PI3K-Akt信号通路等24个子类。结论小鼠肺爆震后早期不同时间点差异表达蛋白组合亦不同,损伤机制复杂,特别是12~24 h肺损伤最为严重,炎症反应显著。Objective To establish the lung blast injury model in mice,detect the proteomic changes of lung in mice at different time points,and explore the mechanism of lung blast injury.Methods A total of 60 healthy male C57BL/6 mice were randomly(random number)divided into the control group,12-h group after thorax blast,24-h group,48-h group,72-h group and 1-week group(n=10 each group).Experiments were carried out in the animal laboratory of the General Hospital of the Northern Theater Command.The model of lung blast injury in mice was established by using a self-developed precision blast device,and the lung tissue injury situation was evaluated by gross observation and HE staining.The proteins in mouse lung tissue were quantitatively analyzed based on LC-MS/MS proteomic technology,and the differentially expressed proteins were screened.On this basis,bioinformatics tool was used to analyze proteomic changes.Results After lung blast injury,scattered bleeding spots could be observed on the surface of lung tissue of mice,and the bleeding points were gradually increased with time,showing a patchy distribution,and the symptoms were the most severe at 24 h.The results of HE staining showed that the normal tissue structure of alveoli disappeared at 12 and 24 h under light microscopy with diffuse bleeding in the alveolar cavity,infiltration of a large number of inflammatory cells,increased interstitial exudate,thickened alveolar wall,and collapsed and merged alveolar cavity.A total of 6861 proteins were identified by LCMS/MS in lung tissue samples of mice after thorax blast,and 608 differentially expressed proteins were quantified,of which 227,140,202,258 and 71 differential proteins were at 12 h,24 h,48 h,72 h,and 1 week,respectively.According to GO analysis,130 biological process subtypes including cell adhesion,extracellular matrix tissue and collagen fibril tissue were obtained.Besides,66 cellular component involving extracellular exosomes,extracellular matrix and cytoplasm were obtained.And 43 molecular functional subclasses

关 键 词：肺爆震伤 急性肺损伤 质谱 蛋白质组学 炎症反应 

分 类 号：R655.3[医药卫生—外科学]