C型产气荚膜梭菌性腹泻仔猪脾脏转录组分析  

Transcriptomic Analysis of the Spleen in Diarrheic Piglet Caused by Clostridium perfringens Type C

作　　者：闫尊强 王鹏飞[1] 杨巧丽 黄晓宇[1] 高小莉[1] 滚双宝[1,2] YAN Zunqiang;WANG Pengfei;YANG Qiaoli;HUANG Xiaoyu;GAO Xiaoli;GUN Shuangbao(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;Gansu Research Center for Swine Production Engineering and Technology,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物科学技术学院,甘肃兰州730070 [2]甘肃省现代养猪技术工程研究中心,甘肃兰州730070

出　　处：《华北农学报》2023年第5期188-196,共9页Acta Agriculturae Boreali-Sinica

基　　金：甘肃农业大学畜牧学养猪产业技术创新团队项目(GAU-XKTD-2022-25);甘肃省教育科技创新项目(GSSYLXM-02);鲁甘科技协作项目(YDZX2021113);2022年现代丝路寒旱农业科技支撑项目(GSLK-2022-10);甘肃农业大学人才专项(GAU-KYQD-2019-07)。

摘　　要：旨在获得C型产气荚膜梭菌性腹泻仔猪血清炎症因子含量、脾脏病理变化及转录组表达谱特征,为其防治提供参考。选择健康状况良好、体质量相近的7日龄二元仔猪6头,随机分为2组,即对照组(CS)和处理组(TS)。TS组每头仔猪每天灌服1 mL菌液(1×109 cfu/mL),连续5 d,CS组仔猪灌服不含菌液的培养液,感染后0,1,3,5 d采集血液分离血清,用ELISA方法检测IL-8、IFN-α和IFN-γ含量。试验结束后屠宰仔猪采集脾脏,进行HE染色和RNA-Seq测序,对测序获得的差异表达基因进行GO功能和KEGG信号通路富集分析,并采用RT-qPCR检测仔猪感染C型产气荚膜梭菌过程中关键差异表达基因的表达量。结果表明:TS组仔猪脾脏出现明显炎症,中性粒细胞浸润,伴有变性及坏死的实质细胞;第3,5天CS组血清中IL-8、IFN-α和IFN-γ含量极显著低于(P<0.01)TS组;TS组与CS组间有1079个差异表达基因,上调583个,下调496个;差异表达基因富集的GO功能主要有细胞对β干扰素的反应、细胞对DNA损伤刺激的反应、炎症反应的负调控等,富集的KEGG信号通路主要有肺结核、乙型肝炎、B细胞受体等;抵抗C型产气荚膜梭菌感染的候选基因(如JAK3、TLR3、ILF2、CXXC1等)RNA-Seq结果与RT-qPCR结果一致。以上结果表明,JAK3、TLR3等基因通过关键信号通路可能在仔猪脾脏抵抗C型产气荚膜梭菌感染过程中发挥了重要功能。The aim of this study was to obtain the serum inflammatory factor content,spleen pathological changes and transcriptome expression profile characteristics of diarrheic piglets with Clostridium perfringens type C,and to provide a reference for the diarrhea prevention and treatment.Six 7-day-old binary piglets with good health and similar body weight were randomly selected and divided into two groups,namely control group(CS)and treatment group(TS).Each piglet in the TS group was fed with 1 mL bacterial solution(1×109 cfu/mL)every day for 5 consecutive days.Then,piglets in the CS group were fed with culture medium without the bacterial solution.Blood samples were collected and serum was separated on 0,1,3 and 5 days after infection.The serum IL-8,IFN-αand IFN-γconcentration were detected by ELISA.After the experiment,piglets were slaughtered to collect the spleen.The partial spleen was collected for HE staining to observe the pathological changes.Additionally,differentially expressed(DE)genes in the spleen were obtained using RNA-Seq.GO terms and KEGG signaling pathways enrichment analysis were performed.The key spleen genes during the infection of piglets with C.perfringens type C were verified by RT-qPCR.The spleen of piglets in the TS group displayed obvious inflammation,neutrophil infiltration,accompanied by degeneration and necrosis of parenchymal cells;on the 3rd and 5th day,the levels of IL-8,IFN-α,and IFN-γin the CS group serum were extremely significantly lower than those in the TS group(P<0.01);a total of 1079 DG genes was screened between TS group and CS group,including 583 up-regulated genes and 496 down-regulated genes.DG genes were mainly enriched in GO terms such as cell response to interferon-β,cell response to DNA damage stimulation,and negative regulation of inflammatory response.Differentially expressed gene were mainly enriched in KEGG signaling pathways such as tuberculosis,hepatitis B,and B cell receptors.These GO terms and KEGG signaling pathways were involved in piglet resistance to C.p

关 键 词：仔猪 腹泻 脾脏 C型产气荚膜梭菌 转录组 

分 类 号：S828[农业科学—畜牧学]