基于蛋白芯片技术对猪附红细胞体感染小鼠血清差异蛋白的筛选  

Screening of serum differential proteins in mice infected with Mycoplasma suis based on protein chip technology

作　　者：梁喜植 于天 王洋[1] 相思宇 闫宗斌 李文学 李海峰[1] 薛书江[1] LIANG Xizhi;YU Tian;WANG Yang;XIANG Siyu;YAN Zongbin;LI Wenxue;LI Haifeng;XUE Shujiang(Department of Veterinary Medicine,Agriculture College of Yanbian University,Yanji,Jilin 133002,China)

机构地区：[1]延边大学、农学院、动物医学系,吉林延吉133002

出　　处：《中国兽医学报》2023年第1期185-190,共6页Chinese Journal of Veterinary Science

基　　金：吉林省自然科学基金资助项目(YDZJ202201ZYTS616);吉林省教育厅“十三五”科学技术资助项目(JJKH20191134KJ)。

摘　　要：为了筛选猪附红细胞体感染小鼠血清的差异蛋白,探讨猪附红细胞体病的致病机制。本试验利用分离纯化后的猪附红细胞体感染昆明种小鼠,设立空白对照组、阴性对照组。经PCR鉴定感染成功后,采集感染小鼠和未感染小鼠血清进行蛋白芯片检测,获取差异蛋白并进行层次聚类分析、功能聚集分析以及KEGG通路富集分析,并对筛选出的细胞因子进行ELISA定量检测验证。结果显示,蛋白芯片检测共筛选出5个差异蛋白,分别是白介素1α(IL-1α)、白介素2(IL-2)、血管内皮生长因子(VEGF)、催乳素Prolactin和神经纤毛蛋白1(Neuropilin-1)。KEGG通路富集分析结果显示:5种差异蛋白主要参与细胞因子与细胞因子受体的互作通路、JAK-STAT信号通路、PI3K-Akt信号通路以及MAPK信号通路。ELISA定量检测结果显示,筛选出的5个差异蛋白含量均显著上升(P<0.05),与蛋白芯片试验结果一致。猪附红细胞体感染可引起小鼠血清中的IL-1α、IL-2、VEGF、Prolactin和Neurpilin-1细胞因子水平显著升高。The aim of this study is to screen the differential proteins in serum of mice infected with Mycoplasma suis and explore the pathogenic mechanism of Mycoplasma suis.Kunming mice were infected with purified Mycoplasma suis,and the blank control group and the negative control group were established.After the successful infection was identified by PCR,the serum of infected and non-infected mice was collected for protein chip detection,and the differential proteins were obtained.The hierarchical cluster analysis,functional aggregation analysis and KEGG pathway enrichment analysis were performed,and the screened cytokines were verified by ELISA quantitative detection.The results showed that a total of five differential proteins were screened out by protein chip detection,namely,interleukin 1α,interleukin 2,vascular endothelial growth factor,prolactin and neurocillin 1.KEGG pathway enrichment analysis showed that the five differentially expressed proteins were mainly involved in the interaction between cytokines and cytokine receptors,JAK-STAT signaling pathway,PI3K-Akt signaling pathway and MAPK signaling pathway.The results of ELISA showed that the five differentially expressed proteins were significantly increased(P<0.05),which was consistent with the results of protein chip test.The levels of IL-1α,IL-2,VEGF,prolactin and neurpilin-1 cytokines in mice serum were significantly increased by Mycoplasma suis infection.

关 键 词：猪附红细胞体 蛋白芯片 差异蛋白 细胞因子 

分 类 号：S852.4[农业科学—基础兽医学]