C型产气荚膜梭菌感染鹿肠道中关键基因和途径的转录组分析

doi:10.11843/j.issn.0366-6964.2023.05.035

摘要/Abstract

摘要： 鹿的产气荚膜梭菌(Clostridium perfringens, C. perfringens)病主要表现为肠毒血症,致死率高。本研究旨在探究C型产气荚膜梭菌感染鹿肠道的关键基因和途径。通过构建C型产气荚膜梭菌感染鹿空肠结扎环模型,HE染色检测C型产气荚膜梭菌感染后空肠的病理学变化,采用Illumina NovaSeq对C型产气荚膜梭菌感染鹿肠道进行转录组测序,并利用qPCR技术进一步验证部分差异表达基因(differentially expressed genes,DEGs);最后通过STRING网站预测多基因蛋白互作网络。结果显示:形态学分析显示,产气荚膜梭菌感染后鹿肠道组织出现明显的坏死和出血。转录组分析显示,一共存在705个差异表达基因,其中,在感染组肠道中有276个上调的差异基因,有429个下调的差异基因。GO富集分析显示,DEGs主要富集在生物调节、对刺激的反应和免疫系统过程;KEGG富集分析显示,DEGs富集的通路主要是T细胞受体信号传导途径、Toll样受体信号通路和造血细胞谱系等,造成肠道损伤和出血,促进鹿肠毒血症的进程。此外,qPCR结果证实了分析结果准确可信。DEGs蛋白互作分析结果表明TLR6、CLDN1、CD3E、IL1A和CCL20与其他蛋白有较多互作关系,并且GO分析和KEGG分析显著富集的通路高度重合。其中,TLR6可能通过抑制紧密连接蛋白表达和免疫炎症反应在鹿产气荚膜梭菌诱导的肠毒血症中发挥关键作用。综上所述,C型产气荚膜梭菌通过破坏细胞膜,并抑制紧密连接蛋白表达,诱导免疫炎症反应,造成肠道出血和损伤。本研究为鹿感染产气荚膜梭菌后调控的分子机制提供了新的思路,为该病的治疗提供理论基础。

关键词: C型产气荚膜梭菌, 肠毒血症, 空肠结扎环模型, 鹿, 转录组分析

Abstract: Clostridium perfringens (C. perfringens) disease in deer mainly manifests as enterotoxemia with high lethality. The aim of this study was to investigate the key genes and pathways of C. perfringens type C infection in the intestine of deer. A C. perfringens-infected deer jejunum ligation loop model was constructed, HE staining was used to detect pathological changes in the jejunum after C. perfringens infection, Illumina NovaSeq was used to sequence the transcriptome of C. perfringens-infected deer intestine, and qPCR was used to further validate some differentially expressed genes (differentially expressed genes (DEGs); finally, the multigene protein interaction network was predicted by STRING website. Morphological analysis revealed significant necrosis and hemorrhage in deer intestinal tissues following C. perfringens infection. Transcriptome analysis indicated the presence of a total of 705 DEGs, of which 276 DEGs up-regulated and 429 DEGs down-regulated genes were present in the infected group intestine. GO enrichment analysis showed that DEGs was mainly enriched in biological regulation, response to stimulus and immune system process; KEGG enrichment analysis showed that the pathways enriched in DEGs were mainly T cell receptor signaling pathways, Toll-like receptor signaling pathway and hematopoietic cell lineage, which caused intestinal damage and bleeding, and promoted the progression of deer enterotoxemia. In addition, the qPCR results confirmed that the analysis was accurate and reliable. The results of protein interaction analysis of DEGs showed that TLR6, CLDN1, CD3E, IL1A and CCL20 had more interactions with other proteins, and the pathways significantly enriched by GO analysis and KEGG analysis were highly overlapping. Among them, TLR6 may play a key role in C. perfringens-induced enterotoxemia in deer through inhibition of tight junction protein expression and immunoinflammation response. C. perfringens type C induced immunoinflammation response by disrupting cell membranes and inhibiting tight junction protein expression, caused intestinal bleeding and intestinal injury. This study provided new ideas on the molecular mechanism of regulation after C. perfringens infection in deer and provided a theoretical basis for the treatment of the disease.

Key words: Clostridium perfringens type C, intestinal loop model, enterotoxemia, deer, transcriptome

中图分类号:

S858.255.1

王美慧, 钟震宇, 白加德, 单云芳, 程志斌, 张庆勋, 孟玉萍, 董玉兰, 郭青云. C型产气荚膜梭菌感染鹿肠道中关键基因和途径的转录组分析[J]. 畜牧兽医学报, 2023, 54(5): 2147-2157.

WANG Meihui, ZHONG Zhenyu, BAI Jiade, SHAN Yunfang, CHENG Zhibin, ZHANG Qingxun, MENG Yuping, DONG Yulan, GUO Qingyun. Transcriptomic Analysis of Key Genes and Pathways in Deer Gut Infected by Clostridium perfringens Type C[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2147-2157.

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