转录组学联合蛋白质组学测序探索与ARDS肺组织病理变化相关的血清生物标志物  

Exploration of serum biomarkers associated with pathological changes in acute respiratory distress syndrome lung tissues through transcriptomics and proteomics

作　　者：龚睿 王乐 龙刚宇 汪茜[1] 姚尚龙[3] 尚游[1] 张定宇 Gong Rui;Wang Le;Long Gangyu;Wang Qian;Yao Shanglong;Shang You;Zhang Dingyu(Department of Critical Care Medicine,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430021,China;Department of Critical Care Medicine,People's Liberation Army 95829 Military Hospital,Wuhan 430014,China;Department of Anesthesiology,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430021,China)

机构地区：[1]华中科技大学同济医学院附属协和医院重症医学科,武汉430021 [2]中国人民解放军95829部队医院重症医学科,武汉430014 [3]华中科技大学同济医学院附属协和医院麻醉科,武汉430021

出　　处：《国际麻醉学与复苏杂志》2024年第11期1141-1150,共10页International Journal of Anesthesiology and Resuscitation

基　　金：湖北省科技创新计划重点研发项目(2023BCB091)。

摘　　要：目的探索与肺源性急性呼吸窘迫综合征(ARDS)肺组织病理蛋白变化一致的血清蛋白标志物。方法采用随机数字表法将40只SPF级健康C57BL/6小鼠分为2组:对照组(Control组,19只)和ARDS实验组[内毒素(LPS)组,21只]。LPS组小鼠给予LPS气道注射,Control组小鼠经气道注射等体积的生理盐水。模型构建后24h,将两组小鼠麻醉后处死,取肺组织进行苏木精-伊红(H-E)染色和湿/干重比(W/D)检测评估ARDS疾病模型;另外对Control组和LPS组小鼠肺组织分别进行转录组学和蛋白质组学测序,并对两组小鼠的血清进行蛋白质组学测序,然后将上述测序数据与本课题组前期关于新型冠状病毒感染(COVID-19)导致的ARDS患者的血清和肺组织蛋白质组学数据进行联合分析,以筛选与肺组织病理蛋白变化一致的血清蛋白标志物。结果与Control组比较,LPS组小鼠肺组织病理损伤评分和肺W/D均升高(均P<0.05),ARDS小鼠疾病模型构建成功。通过对小鼠肺组织转录组学和蛋白质组学测序以及血清蛋白质组学测序,共鉴定出28个共同的差异表达蛋白(CM-DEP),这些蛋白在ARDS小鼠肺组织及血清中的变化趋势一致。进一步结合ARDS患者肺组织与血浆蛋白质组学测序结果,筛选出4个候选蛋白,即结合珠蛋白(HP)、S100钙结合蛋白A9(S100A9)、血清淀粉样蛋白A1(SAA1)、血清淀粉样蛋白A2(SAA2),它们与ARDS患者病情严重程度呈正相关。结论4个候选蛋白(HP、S100A9、SAA1和SAA2)有潜力成为反映ARDS肺组织病理变化的血清生物标志物,且可能具有良好的临床价值。Objective To explore serum protein biomarkers that align with the pathological protein changes in lung tissues of pulmonary acute respiratory distress syndrome(ARDS).Methods A total of 40 healthy SPF-grade C57BL/6 mice were divided into two groups according to the random number table method:a control group(Control group,n=19)and an ARDS experimental group[lipo-polysaccharide(LPS)group,n=21].The LPS group was administered with LPS through intratracheal injection,while the Control group received an equal volume of saline through intratracheal injection.Twenty-four hours after modeling,mice in both groups were euthanized under anesthesia.Lung tissues were collected for hematoxylin-eosin(H-E)staining and wet/dry(W/D)analysis to evaluate the ARDS model.In addition,lung tissues from the Control and LPS groups were collected for transcriptomic and proteomic sequencing.Serum samples from both groups were also subjected to proteomic analysis.These sequencing results were then integrated with previous proteomic data from serum and lung tissues of coronavirus disease 2019(COVID-19)induced ARDS patients obtained by the research team,in an effort to identify serum protein biomarkers consistent with the pathological protein changes in lung tissues.Results Compared with the Control group,the LPS group showed increases in pathological injury scores in lung tissues and pulmonary W/D(all P<0.05),and the ARDS model of mice was successfully constructed.A total of 28 common differentially expressed proteins(CM-DEP)were identified through transcriptomic and proteomic analysis of lung tissues,along with proteomic sequencing of serum.These proteins exhibited consistent trends in both lung tissues and serum of ARDS mice.Further analysis incorporating proteomic data from lung tissues and plasma of ARDS patients identified four candidate proteins,namely haptoglobin(HP),S100 calcium-binding protein A9(S100A9),serum amyloid A1(SAA1),and serum amyloid A2(SAA2).These proteins showed a positive correlation with the severity of ARDS in patients

关 键 词：急性呼吸窘迫综合征 血清 生物标志物 转录组学 蛋白质组学 

分 类 号：R563.8[医药卫生—呼吸系统]