C型产气荚膜梭菌外毒素致小鼠肠道损伤的转录组分析

Transcriptome Analysis of Intestinal Injury Induced by Clostridium perfringens Type C Exotoxin in Mouse

旨在探索产气荚膜梭菌外毒素造成机体炎性损伤及免疫调控紊乱的毒性机制,为产气荚膜梭菌病的致病机理研究提供理论基础。将C型产气荚膜梭菌强毒株C59-2培养上清腹腔注射BALB/c小鼠,采集小肠样本进行转录组测序,筛选差异表达基因,并对其进行GO功能注释和KEGG通路富集分析。结果显示,共获得40.99 Gb有效碱基,筛选后共得到795个差异表达基因,其中,229个基因表达上调,566个基因表达下调,对随机选取的10个基因进行q-PCR验证,其相对表达量与转录表达谱一致。GO功能注释主要涉及G蛋白偶联核苷酸受体活性和G蛋白偶联嘌呤核苷酸受体活性等。KEGG通路富集分析发现,主要富集在TNF信号通路、IL-17信号通路、p53信号通路、FOXO信号通路、Toll样受体信号通路、NF-κB信号通路等。产气荚膜梭菌外泌毒素侵入机体,会激活TNF等炎性信号通路,进而造成肠道发生炎性损伤甚至坏死。

The present study aimed to explore the toxicity mechanism of inflammatory injury and immune regulation disorders in organisms,and lay a foundation for further in-depth exploration of the pathogenic mechanism of Clostridium perfringens disease.The cultured supernatant of C.perfringens type C was intraperitoneally injected into BALB/c mice,and the small intestine samples were collected for transcriptome sequencing.Based on the differentially expressed genes(DEGs),GO functional annotation and KEGG pathway enrichment analysis were performed.The results showed that in total,there were 40.99 Gb effective bases and 795 DEGs were obtained,among which 229 were up-regulated and 566 were down-regulated.The ten randomly selected DEGs were verified by real-time fluorescent quantitative PCR that their relative expression was consistent with the transcriptional expression profile.GO functional annotation mainly involved G-protein coupled nucleotide receptor activity,and G-protein coupled purine nucleotide receptor activity.KEGG pathway enrichment analysis found that it was mainly concentrated in TNF signaling pathway,IL-17 signaling pathway,p53 signaling pathway,FOXO signaling pathway,Toll-like receptor signaling pathway,and NF-κB signaling pathway.When C.perfringens exotoxin invades the body,inflammatory signaling pathways such as TNF will be activated,resulting in inflammatory damage and even necrosis of the intestine.