细粒棘球蚴囊液蛋白组分分析  

Analysis of components of proteins from Echinococcus granulosus cyst fluid

作　　者：王舒颖 杨小迪 高宏宇[1,2] 邢敬雨 胡琪 黄婷婷 吴培 赵艺婷 刘浩伟 刘为玥 王会娜 周瑞 褚亮[4] WANG Shu-Ying;YANG Xiao-Di;GAO Hong-Yu;XING Jing-Yu;HU Qi;HUANG Ting-Ting;WU Pei;ZHAO Yi-Ting;LIU Hao-Wei;LIU Wei-Yue;WANG Hui-Na;ZHOU Rui;CHU Liang(Department of Pediatrics,The First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China;Anhui Key Laboratory of Infection and Immunity,Bengbu Medical College,China;Department of Microbiology and Parasitology,Bengbu Medical College,China;Department of General Surgery,The Second Affiliated Hospital of Bengbu Medical College,China)

机构地区：[1]蚌埠医学院第一附属医院儿科,蚌埠233000 [2]安徽省感染与免疫重点实验室 [3]蚌埠医学院病原生物学教研室 [4]蚌埠医学院第二附属医院普外科

出　　处：《中国血吸虫病防治杂志》2021年第5期476-482,539,共8页Chinese Journal of Schistosomiasis Control

基　　金：安徽省高校学科(专业)拔尖人才学术资助项目(gxbj ZD15);安徽省卫生健康委科研重点项目(AHWJ2021a005);安徽省教育厅重点自然科学研究项目(KJ2019A0383);蚌埠医学院自然科学高层次科技项目立项攀登计划项目(2020bypd007);蚌埠医学院“512人才培育计划”(by51201205);蚌埠医学院研究生科研创新计划项目(Byy-cxz20015);蚌埠市-蚌埠医学院联合科技攻关项目(BYLK201818);国家级大学生创新训练项目(202010367006、201910367002)。

摘　　要：目的利用鸟枪法分析细粒棘球蚴(Echinococcus granulosus)囊液蛋白(hydatid cyst fluid protein,HCFP)组分,寻找其中具有潜在调控免疫失调性疾病功能的活性组分。方法无菌收集细粒棘球蚴病患者肝囊肿中的细粒棘球蚴囊液,采用鸟枪法进行液相色谱-串联质谱鉴定,采用Max Quant 1.6.1.0软件进行数据库检索。利用基因本体论(Gene Ontol-ogy,GO)对已鉴定的蛋白质从细胞组分、分子功能和生物学功能三个方面进行分类。结果经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离的细粒棘球蚴囊液蛋白条带相对分子量为25~70 k Da,质谱分析共获得37个蛋白,其中已鉴定蛋白32种、未知蛋白5种。已初步发现至少有4种蛋白具有潜在调节免疫失调性疾病作用,即抗原B、谷胱甘肽S-转移酶、硫氧还蛋白过氧化物酶和苹果酸脱氢酶。GO富集分析显示,已鉴定蛋白具有149种分子功能,参与341种生物学过程。结论细粒棘球蚴囊液蛋白成分复杂多样,从已知蛋白中初步筛选出4种与调节免疫失调性疾病潜在相关的蛋白。Objective To analyze the components of proteins from Echinococcus granulosus cyst fluid using the shotgun meth-od, and to identify the active components with potential regulatory effects for immune dysregulation diseases.Methods TheE. granulosus cyst fluid was collected aseptically from the hepatic cysts of patients with cystic echinococcosis, and characterizedby liquid chromatography(LC)tandem mass spectrometry(MS/MS)following digestion with trypsin. The protein data weresearched using the software Max Quant version 1.6.1.0 and the cellular components, molecular functions, and biological processesof the identified proteins were analyzed using the Gene Ontology(GO)method.Results The E. granulosus cyst fluid separatedby sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)had a relative molecular mass of 25 to 70 k Da. LS-MS/MS analysis identified 37 proteins, including 32 known proteins and 5 unknown proteins. At least 4 proteins were preliminari-ly found to exhibit potential regulatory effects for immune dysregulation diseases, including antigen B, glutathione-S-transferase(GST), thioredoxin peroxidase(TPX)and malate dehydrogenase(MDH). GO enrichment analysis showed that the identified proteins had 149 molecular functions and were involved in 341 biological processes.Conclusions E. granulosuscyst fluid hasa variety of protein components, and four known proteins are preliminarily identified to be associated with immune dysregulationdiseases.

关 键 词：细粒棘球蚴 囊液 蛋白质组 鸟枪法 免疫失调性疾病 

分 类 号：R383.33[医药卫生—医学寄生虫学]