基于数据非依赖型采集蛋白质组学分析劳力性热射病大鼠血浆蛋白的变化  

The changes of plasma protein in the rats with exertional heat stroke by DIA proteomics

作　　者：林青伟 罗竹青 何龙平 余永春 曾俊杰 宋景春[1] Lin Qingwei;Luo Zhuqing;He Longping;Yu Yongchun;Zeng Junjie;Song Jingchun(Department of Critical Care Medicine,No.908th Hospital of Joint Logistic Support Force of Chinese PLA,Nanchang 330002,China)

机构地区：[1]中国人民解放军联勤保障部队第九〇八医院重症医学科、南昌市血栓与止血学重点实验室,江西南昌330002

出　　处：《中国急救医学》2024年第6期495-502,共8页Chinese Journal of Critical Care Medicine

基　　金：2022年全军基础加强计划项目(2022-JCJQ-ZD-097-11);中国医药教育学会2022科学攻关科研课题(2022KTZ013)。

摘　　要：目的采用数据非依赖型采集(DIA)蛋白质组学技术分析劳力性热射病(EHS)大鼠血浆蛋白表达的变化。方法取SPF级雄性SD大鼠10只,随机分为正常对照组(CON组,n=5)和EHS组(n=5),EHS组造模前1周完成温度遥测胶囊植入术,术毕恢复7 d后大鼠在人工气候舱内(温度40℃,相对湿度70%)跑步,监测核心温度达到42.5℃时视为发生EHS。停止跑步后采血进行DIA定量蛋白质组学分析。结果以表达倍数>1.5倍且P<0.05为筛选标准,共筛选出625个上调差异表达蛋白(DEPs)和8个下调DEPs。亚细胞定位注释表明,DEPs主要位于细胞外和细胞质;基因本体论(GO)注释显示,DEPs分子主要功能为蛋白质结合;京都基因与基因组百科全书(KEGG)注释显示,DEPs主要参与免疫反应、能量代谢等信号通路;同源蛋白簇(COG)注释显示,DEPs主要发挥翻译后修饰、蛋白质周转和伴侣功能。蛋白结构域富集分析显示,DEPs结构域主要集中在免疫球蛋白结构域、血小板反应蛋白1型结构域和补体Clr样EGF样结构域;GO富集分析显示,DEPs主要参与血液凝固和纤维蛋白凝块形成、纤溶酶原激活、急性炎症反应的调节等过程;KEGG富集分析显示,DEPs主要参与补体和凝血级联、免疫和细胞黏附分子等信号通路;Reactome通路富集分析显示,DEPs主要参与先天免疫系统、中性粒细胞脱颗粒和血小板活化等信号通路;蛋白互作网络分析发现,参与补体和凝血级联通路的蛋白网络互作较多,且以上调蛋白Fgg、Plg和Serpinc1为主。结论EHS明显改变大鼠血浆中蛋白表达谱,主要涉及免疫系统异常激活、炎症反应失控和凝血功能紊乱的信号通路。Objective To analyze the changes of plasma protein expression in the rats with exertional heat stroke(EHS)by using data-independent acquisition(DIA)proteomics.Methods Ten specific pathogen-free(SPF)male SD rats were randomly divided into a normal control group(n=5)and an EHS group(n=5).The EHS group underwent temperature telemetry capsule implantation surgery 1 week before modeling,and after one week of recovery,the rats ran in an artificial climate chamber(temperature 40℃,relative humidity 70%)until the core temperature reached 42.5℃,which was considered the occurrence of EHS.Blood was collected after running to analyze the DIA quantitative proteomics.Results A total of 625 upregulated differentially expressed proteins(DEPs)and 8 downregulated DEPs were screened by using the criteria of expression multiple>1.5 times and P<0.05.Subcellular localization annotation showed that DEPs were mainly located in the extracellular space and cytoplasm.Gene ontology(GO)annotation showed that DEPs mainly functioned as protein binding.Kyoto encyclopedia of genes and genomes(KEGG)annotation showed that DEPs mainly participated in immune response,energy metabolism,and other signaling pathways.Clusters of orthologous groups of proteins(COG)annotation showed that DEPs mainly played roles in post-translational modification,protein turnover and chaperone function.Protein domain enrichment analysis showed that the protein domains of DEPs were mainly concentrated in immunoglobulin domains,platelet reactivity protein 1 type domains,and complement Clr-like EGF-like domains.GO enrichment analysis showed that DEPs mainly participated in blood coagulation and fibrin clot formation,fibrinolytic enzyme activation,and acute inflammatory response regulation.KEGG enrichment analysis showed that DEPs mainly participated in the complement and coagulation cascades,immune and cell adhesion molecule signaling pathways.Reactome pathway enrichment analysis showed that DEPs mainly participated in the innate immune system,platelet activation,and neut

关 键 词：热射病 蛋白质组学 数据非依赖型采集 凝血 大鼠 

分 类 号：R594.12[医药卫生—内科学]