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

Abstract

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

CLC Number:

S858.255.1

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.

References 31

[1]	OHTANI K, SHIMIZU T. Regulation of toxin production in Clostridium perfringens[J]. Toxins (Basel), 2016, 8(7):207.
[2]	ROOD J I, ADAMS V, LACEY J, et al. Expansion of the Clostridium perfringens toxin-based typing scheme[J]. Anaerobe, 2018, 53:5-10.
[3]	HASSAN K A, ELBOURNE L D H, TETU S G, et al. Genomic analyses of Clostridium perfringens isolates from five toxinotypes[J]. Res Microbiol, 2015, 166(4):255-263.
[4]	MVLLER K E, ROZGONYI F. Élelmiszer eredetü bakteriális megbetegedések patogenezise, klinikai jellegzetességei, diagnosztikája és kezelése[J]. Orv Hetil, 2020, 161(48):2019-2028.
[5]	闫新华, 赵传芳, 王长凤, 等. 鹿肠毒血症的综合诊断[J]. 特种经济动植物, 2002, 5(10):38-39.YAN X H, ZHAO C F, WANG C F, et al. Comprehensive diagnosis of deer enterotoxemia[J]. Special Economic Animal and Plant, 2002, 5(10):38-39. (in Chinese)
[6]	朱凯宗. 山东地区鹿肠毒血症的综合诊断[J]. 畜牧兽医科技信息, 2019(9):169-170.ZHU K Z. Comprehensive diagnosis of deer intestine toxemia in Shandong area[J]. Chinese Journal of Animal Husbandry and Veterinary Medicine, 2019(9):169-170. (in Chinese)
[7]	张成林, 刘燕, 闫鹤, 等. 麋鹿发生C型魏氏梭菌病[J]. 野生动物, 2012, 33(5):260-263.ZHANG C L, LIU Y, YAN H, et al. C-type clostridia gastroenteritis in Pere David's deer Elaphurus davidianus[J]. Chinese Journal of Wildlife, 2012, 33(5):260-263. (in Chinese)
[8]	UZAL F A, MCCLANE B A. Recent progress in understanding the pathogenesis of Clostridium perfringens type C infections[J]. Vet Microbiol, 2011, 153(1-2):37-43.
[9]	ROOD J I, COLE S T. Molecular genetics and pathogenesis of Clostridium perfringens[J]. Microbiol Rev, 1991, 55(4):621-648.
[10]	ODA M, TERAO Y, SAKURAI J, et al. Membrane-binding mechanism of Clostridium perfringens alpha-toxin[J]. Toxins (Basel), 2015, 7(12):5268-5275.
[11]	VAN ASTEN A J A M, NIKOLAOU G N, GRÖNE A. The occurrence of cpb 2-toxigenic Clostridium perfringens and the possible role of the β2-toxin in enteric disease of domestic animals, wild animals and humans[J]. Vet J, 2010, 183(2):135-140.
[12]	ZENG X, LIU B S, ZHOU J, et al. Complete genomic sequence and analysis of β2 toxin gene mapping of Clostridium perfringens JXJA17 isolated from piglets in China[J]. Sci Rep, 2021, 11(1):475.
[13]	祁云霞, 刘永斌, 荣威恒. 转录组研究新技术:RNA-Seq及其应用[J]. 遗传, 2011, 33(11):1191-1202.QI Y X, LIU Y B, RONG W H. RNA-Seq and its applications:a new technology for transcriptomics[J]. Hereditas (Beijing), 2011, 33(11):1191-1202. (in Chinese)
[14]	BROOKS A N, TURKARSLAN S, BEER K D, et al. Adaptation of cells to new environments[J]. Wiley Interdiscip Rev Syst Biol Med, 2011, 3(5):544-561.
[15]	SONGER J G. Clostridial diseases of small ruminants[J]. Vet Res, 1998, 29(3-4):219-232.
[16]	SONGER J G, UZAL F A. Clostridial enteric infections in pigs[J]. J Vet Diagn Invest, 2005, 17(6):528-536.
[17]	MEHDIZADEH GOHARI I, NAVARRO M A, LI J H, et al. Pathogenicity and virulence of Clostridium perfringens[J]. Virulence, 2021, 12(1):723-753.
[18]	UZAL F A, VIDAL J E, MCCLANE B A, et al. Clostridium perfringens toxins involved in mammalian veterinary diseases[J]. Open Toxinol J, 2010, 2:24-42.
[19]	SHI H R, HUANG X Y, YAN Z Q, et al. Effect of Clostridium perfringens type C on TLR4/MyD88/NF-κB signaling pathway in piglet small intestines[J]. Microb Pathog, 2019, 135:103567.
[20]	LEPP D, ZHOU Y, OJHA S, et al. Clostridium perfringens produces an adhesive pilus required for the pathogenesis of necrotic enteritis in poultry[J]. J Bacteriol, 2021, 203(7):e00578-20.
[21]	POPOFF M R. Clostridial pore-forming toxins:powerful virulence factors[J]. Anaerobe, 2014, 30:220-238.
[22]	NAGAHAMA M, OCHI S, ODA M, et al. Recent insights into Clostridium perfringens beta-toxin[J]. Toxins (Basel), 2015, 7(2):396-406.
[23]	KIMURA J, ABE H, KAMITANI S, et al. Clostridium perfringens enterotoxin interacts with claudins via electrostatic attraction[J]. J Biol Chem, 2010, 285(1):401-408.
[24]	LU M M, YUAN B H, YAN X H, et al. Clostridium perfringens -induced host-pathogen transcriptional changes in the small intestine of broiler chickens[J]. Pathogens, 2021, 10(12):1607.
[25]	ABREU M T. Toll-like receptor signalling in the intestinal epithelium:how bacterial recognition shapes intestinal function[J]. Nat Rev Immunol, 2010, 10(2):131-144.
[26]	KAUR A, KAUSHIK D, PIPLANI S, et al. TLR2 agonistic small molecules:detailed Structure-activity relationship, applications, and future prospects[J]. J Med Chem, 2021, 64(1):233-278.
[27]	NOREEN M, ARSHAD M. Association of TLR1, TLR2, TLR4, TLR6, and TIRAP polymorphisms with disease susceptibility[J]. Immunol Res, 2015, 62(2):234-252.
[28]	DU E C, WANG W W, GAN L, et al. Effects of thymol and carvacrol supplementation on intestinal integrity and immune responses of broiler chickens challenged with Clostridium perfringens[J]. J Anim Sci Biotechnol, 2016, 7:19.
[29]	TAKEDA K, AKIRA S. TLR signaling pathways[J]. Semin Immunol, 2004, 16(1):3-9.
[30]	LUO R R, YANG Q L, HUANG X Y, et al. Clostridium perfringens beta2 toxin induced in vitro oxidative damage and its toxic assessment in porcine small intestinal epithelial cell lines[J]. Gene, 2020, 759:144999.
[31]	HUANG F C. The interleukins orchestrate mucosal immune responses to Salmonella infection in the intestine[J]. Cells, 2021, 10(12):3492.

Transcriptomic Analysis of Key Genes and Pathways in Deer Gut Infected by Clostridium perfringens Type C

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