早期皮下寄生多房棘球蚴的蛋白组学分析  

Proteomic analysis of early subcutaneous parasitic Echinococcus multilocularis in mice

作　　者：兰天艳 张永娥 郭鹏辉[1] 郭小腊 陈轶霞[1] 王明明[1] 郑亚东[2] LAN Tian-yan;ZHANG Yong-e;GUO Peng-hui;GUO Xiao-la;CHEN Yi-xia;WANG Ming-ming;ZHENG Ya-dong(College of Life Science and Engineering,Northwest Minzu University,Lanzhou 730030,China;State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]西北民族大学生命科学与工程学院,兰州730030 [2]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室/甘肃省动物寄生虫病重点实验室,兰州730046

出　　处：《中国人兽共患病学报》2020年第3期187-193,共7页Chinese Journal of Zoonoses

基　　金：西北民族大学中央高校基本科研业务费项目(No.31920190019,31920190202-03,No.31920190027);国家自然科学基金项目(No.U1703104,No.31472185);国家重点基础研究发展计划(973计划)项目(No.2015CB150300)联合资助。

摘　　要：目的了解皮下寄生多房棘球蚴的蛋白质组特征。方法每只BALB/c小鼠皮下多点注射约600只原头蚴,一月后分离皮下寄生的包囊,制备组织切片,HE染色,糖原染色,利用显微镜观察虫体形态,同时提取蛋白质并进行测序、GO基因功能分析和COG分析,最后利用Western blotting对部分蛋白质进行验证。结果皮下寄生的多房棘球蚴呈半透明状,如绿豆大小,包囊内含有囊液。在光学显微镜下,可见包囊中含有子囊,囊中仅有少数原头蚴,且一些原头蚴头节上还带有小钩;囊壁糖原染色阳性,呈粉红色。测序共鉴定203种蛋白质,其中多种蛋白酶的表达水平很高,如磷酸烯醇丙酮酸羧激酶、柠檬酸合酶等。在功能上,所鉴定的蛋白质主要富集在翻译后修饰、蛋白质转换和伴侣分子信号通路、能量合成与转换以及碳水化合物运输与代谢方面,而参与免疫应答、脂质运输与代谢等方面的蛋白质则较少。在丰度较高的前20种蛋白质中,多数为参与糖代谢过程的蛋白质。Western blotting结果显示,VCP、Enolase、14-3-3和β-actin在包囊蛋白中均有不同程度地表达,而GST不表达或表达水平非常低,这与测序结果基本一致。结论研究结果表明,皮下寄生的多房棘球蚴在糖代谢和脂质代谢方面可能发生了明显地改变,这为探索包囊不育的机理提供了有价值的线索。To understand the proteomic characteristics of subcutaneous parasitic Echinococcus multilocularis.Approximately 600 protoscoleces were subcutaneously injected in each BALB/c mice.Subcutaneous parasitic hydatid cysts were isolated after one month.Then the tissue sections were prepared,and HE staining,glycogen staining,and morphological observation by microscope were conducted.Moreover,the proteome of hydatid cysts was determined,and then GO gene function analysis and COG analysis were performed.Finally,some proteins were verified by Western blotting.The subcutaneous parasitic Echinococcus multilocularis was transparent with a size of mung beans,which contained cystic fluid.Under the light microscope,mother capsules contained offspring cysts,and there were only a few protoscoleces,some of which had small hooks.Moreover,the glycogen staining was positive and the cyst wall was in pink.A total of 203 proteins were identified by sequencing,and the expression levels of various proteases were very high,such as phosphoenolpyruvate carboxykinase,citrate synthase and so forth.Functionally,the identified proteins are mainly enriched in post-translational modifications,protein turnover and chaperone signaling pathways,energy synthesis and conversion,and carbohydrate transport and metabolism,while proteins involved in immune responses,lipid transport,and metabolism were less.Among the top 20 proteins with high abundance,most proteins were involved in the process of glucose metabolism.Western blotting results showed that VCP,Enolase,14-3-3 and β-actin were expressed in different degrees in the cyst,while GST was not expressed or expressed at a very low level,which was consistent with the sequencing results.The results suggested that subcutaneous parasitic Echinococcus multilocularis may have significant changes in glucose metabolism and fat metabolism,which provides valuable clues for exploring the mechanism of hydatid cyst infertility.

关 键 词：多房棘球绦虫 包虫病 包囊 蛋白质组学 

分 类 号：R383.3[医药卫生—医学寄生虫学]