Enolase在鸭疫里默氏杆菌侵袭鸭脑微血管内皮细胞中的作用

doi:10.11843/j.issn.0366-6964.2022.12.024

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (12): 4389-4397.doi: 10.11843/j.issn.0366-6964.2022.12.024

Enolase在鸭疫里默氏杆菌侵袭鸭脑微血管内皮细胞中的作用

邹荣华, 吴晓妮, 陈启伟, 宫晓炜, 王燕萍, 郑福英^*, 储岳峰

中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室, 兰州 730046

收稿日期:2021-12-03 出版日期:2022-12-23 发布日期:2022-12-25
通讯作者: 郑福英，主要从事细菌耐药机制和致病机制研究，E-mail：zhengfuying@caas.cn
作者简介:邹荣华(1997-)，女，安徽淮北人，硕士生，主要从事细菌耐药机制和致病机制研究，E-mail：ronghua29@126.com
基金资助:
国家自然科学基金(31572532)；中国农业科学院创新工程经费

Effect of Enolase on Riemerella anatipestifer Invading Duck Brain Microvascular Endothelial Cells

ZOU Ronghua, WU Xiaoni, CHEN Qiwei, GONG Xiaowei, WANG Yanping, ZHENG Fuying^*, CHU Yuefeng

State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2021-12-03 Online:2022-12-23 Published:2022-12-25

摘要/Abstract

摘要： 旨在明确鸭疫里默氏杆菌烯醇化酶(Enolase)在其侵袭鸭脑微血管内皮细胞(DBMEC)以及血脑屏障(BBB)中的作用。本研究以鸭疫里默氏杆菌RA-LZ01株为亲本株，利用同源重组和结合转移的方法构建enolase基因缺失株ΔEnolase和回复株cΔEnolase，并测定RA-LZ01、ΔEnolase和cΔEnolase对DBMEC黏附和侵袭能力的差异；用上述菌株感染雏鸭，测定雏鸭血液和脑组织中的载菌量。结果表明，与亲本株RA-LZ01相比，缺失株ΔEnolase对DBMEC的黏附率和入侵率均极显著降低；回复株cΔEnolase恢复了对DBMEC的黏附和入侵能力。感染RA-LZ01、ΔEnolase和cΔEnolase菌株的雏鸭的血液载菌量无显著差异；与感染RA-LZ01和cΔEnolase菌株的雏鸭相比，感染ΔEnolase菌株的雏鸭脑组织中的载菌量极显著降低。以上结果说明，Enolase与鸭疫里默氏杆菌黏附和入侵DBMEC以及入侵雏鸭脑组织显著相关，可能为介导鸭疫里默氏杆菌突破鸭血脑屏障的毒力因子。

关键词: 鸭疫里默氏杆菌, Enolase, 鸭脑微血管内皮细胞, 血脑屏障

Abstract: This study aimed to clarify the role of Enolase on Riemerella anatipestifer invading duck brain microvascular endothelial cells (DBMEC) and blood brain barrier. R. anatipestifer enolase gene deletion mutant ΔEnolase and the complemented strain cΔEnolase were constructed by homologous recombination and combined transfer assay using RA-LZ01 strain as the parental strain. The differences of adherence and invasion abilities of RA-LZ01, ΔEnolase and cΔEnolase to DBMEC were detected. At the same time, bacterial loads in blood and brain of ducklings infected with RA-LZ01, ΔEnolase and cΔEnolase were determined respectively.The results showed that, compared with RA-LZ01, the adhesion and invasion rates of ΔEnolase to DBMEC were significantly reduced, and cΔEnolase restored the adhesion and invasion abilities to DBMEC. Bacterial loads in blood had no significant difference among the animals infected with RA-LZ01, ΔEnolase or cΔEnolase. Compared with the ducklings infected with RA-LZ01 and cΔEnolase strains, the loads of bacteria in the brain of the ducklings infected with ΔEnolase strain was significantly decreased. These results indicated that Enolase was significantly correlated with the adhesion and invasion of R. anatipestifer into DBMEC and the invasion of ducklings’ brain. Hence, Enolase should be a virulence factor mediating R. anatipestifer crossing duck blood brain barrier.

Key words: Riemerella anatipestifer, Enolase, duck brain microvascular endothelial cells, blood brain barrier

中图分类号:

S858.325.12

邹荣华, 吴晓妮, 陈启伟, 宫晓炜, 王燕萍, 郑福英, 储岳峰. Enolase在鸭疫里默氏杆菌侵袭鸭脑微血管内皮细胞中的作用[J]. 畜牧兽医学报, 2022, 53(12): 4389-4397.

ZOU Ronghua, WU Xiaoni, CHEN Qiwei, GONG Xiaowei, WANG Yanping, ZHENG Fuying, CHU Yuefeng. Effect of Enolase on Riemerella anatipestifer Invading Duck Brain Microvascular Endothelial Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4389-4397.

参考文献 27

[1]	LIU L,JOHNSON H L,COUSENS S,et al.Global,regional,and national causes of child mortality:An updated systematic analysis for 2010 with time trends since 2000[J].Lancet,2012,379(9832):2151-2161.
[2]	DE GANS J,VAN DE BEEK D.Dexamethasone in adults with bacterial meningitis[J].N Engl J Med,2002,347(20):1549-1556.
[3]	VAN DE BEEK D,DE GANS J,SPANJAARD L,et al.Clinical features and prognostic factors in adults with bacterial meningitis[J].N Engl J Med,2004,351(18):1849-1859.
[4]	LI S D,GONG X W,CHEN Q W,et al.Threshold level of Riemerella anatipestifer crossing blood-brain barrier and expression profiles of immune-related proteins in blood and brain tissue from infected ducks[J].Vet Immunol Immunopathol,2018,200:26-31.
[5]	吴彩艳,程淑琴,张建峰,等.我国鸭疫里氏杆菌病流行概述[J].动物医学进展,2017,38(6):86-90.WU C Y,CHENG S Q,ZHANG J F,et al.Introduction to epidemiology of Riemerellosis anatipestifer disease in China[J].Progress in Veterinary Medicine,2017,38(6):86-90.(in Chinese)
[6]	HU Q H,HAN X A,ZHOU X J,et al.OmpA is a virulence factor of Riemerella anatipestifer[J].Vet Microbiol,2011,150(3-4):278-283.
[7]	WENG S C,LIN W H,CHANG Y F,et al.Identification of a virulence-associated protein homolog gene and ISRa1 in a plasmid of Riemerella anatipestifer[J].FEMS Microbiol Lett,1999,179(1):11-19.
[8]	WANG X L,DING C,WANG S H,et al.The AS87_04050 gene is involved in bacterial lipopolysaccharide biosynthesis and pathogenicity of Riemerella anatipestifer[J].PLoS One,2014,9(10):e109962.
[9]	WANG X L,YUE J P,DING C,et al.Deletion of AS87_03730 gene changed the bacterial virulence and gene expression of Riemerella anatipestifer[J].Sci Rep,2016,6:22438.
[10]	ZHAO X X,LIU Q,ZHANG J,et al.Identification of a gene in Riemerella anatipestifer CH-1 (B739-2187) that contributes to resistance to polymyxin B and evaluation of its mutant as a live attenuated vaccine[J].Microb Pathog,2016,91:99-106.
[11]	GUO Y Q,HU D,GUO J,et al.Riemerella anatipestifer type IX secretion system is required for virulence and gelatinase secretion[J].Front Microbiol,2017,8:2553.
[12]	LIU H T,ZHU S,SUN Y Y,et al.Selection of potential virulence factors contributing to Streptococcus suis serotype 2 penetration into the blood-brain barrier in an in vitro Co-culture model[J].J Microbiol Biotechnol,2017,27(1):161-170.
[13]	SUN Y Y,LI N,ZHANG J,et al.Enolase of Streptococcus Suis serotype 2 enhances blood-brain barrier permeability by inducing IL-8 release[J].Inflammation,2016,39(2):718-726.
[14]	XIA X J,QIN W H,ZHU H L,et al.How Streptococcus suis serotype 2 attempts to avoid attack by host immune defenses[J].J Microbiol Immunol Infect,2019,52(4):516-525.
[15]	ZAKRZEWICZ D,BERGMANN S,DIDIASOVA M,et al.Host-derived extracellular RNA promotes adhesion of Streptococcus pneumoniae to endothelial and epithelial cells[J].Sci Rep,2016,6:37758.
[16]	CARNEIRO C R W,POSTOL E,NOMIZO R,et al.Identification of enolase as a laminin-binding protein on the surface of Staphylococcus aureus[J].Microbes Infect,2004,6(6):604-608.
[17]	秦明星,宫晓炜,陈启伟,等.鸭疫里默氏杆菌RA-LZ01株GE296_RS01450基因介导其对重金属离子产生耐受性的分析[J].畜牧兽医学报,2021,52(10):2864-2873.QIN M X,GONG X W,CHEN Q W,et al.Study on the GE296_RS01450 gene-mediated tolerance of Rimerella anatipestifer strain RA-LZ01 to heavy metal ions[J].Acta Veterinaria et Zootechnica Sinica,2021,52(10):2864-2873.(in Chinese)
[18]	权衡,陈启伟,宫晓炜,等.鸭疫里默氏杆菌MFS外排泵rant基因缺失株的构建及其介导的耐药性[J].畜牧兽医学报,2021,52(7):1991-1999.QUAN H,CHEN Q W,GONG X W,et al.Construction of MFS efflux pump rant gene deletion mutant of Riemerella anatipestifer and its contribution to drug resistance[J].Acta Veterinaria et Zootechnica Sinica,2021,52(7):1991-1999.(in Chinese)
[19]	薛亚飞,宫艳杉,祁晶晶,等.鸭疫里默氏杆菌对鸭胚肝细胞粘附入侵能力的研究[J].中国动物传染病学报,2019,27(1):37-41.XUE Y F,GONG Y S,QI J J,et al.Adhesion and invasion of Riemerella anatipestifer to duck embryo liver cells[J].Chinese Journal of Animal Infectious Diseases,2019,27(1):37-41.(in Chinese)
[20]	ZHAO W D,LIU D X,WEI J Y,et al.Caspr1 is a host receptor for meningitis-causing Escherichia coli[J].Nat Commun,2018,9(1):2296.
[21]	TENG C H,CAI M,SHIN S,et al.Escherichia coli K1 RS218 interacts with human brain microvascular endothelial cells via type 1 fimbria bacteria in the fimbriated state[J].Infect Immun,2005,73(5):2923-2931.
[22]	SAUNDERS N R,DREIFUSS J J,DZIEGIELEWSKA K M,et al.The rights and wrongs of blood-brain barrier permeability studies:A walk through 100 years of history[J].Front Neurosci,2014,8:404.
[23]	PANDEY P K,SHARMA A K,GUPTA U.Blood brain barrier:An overview on strategies in drug delivery,realistic in vitro modeling and in vivo live tracking[J].Tissue Barriers,2016,4(1):e1129476.
[24]	KRUCINSKA J,FALCONE E,ERLANDSEN H,et al.Structural and functional studies of bacterial enolase,a potential target against Gram-negative pathogens[J].Biochemistry,2019,58(9):1188-1197.
[25]	ANTIKAINEN J,KUPARINEN V,LÄHTEENMÄKI K,et al.Enolases from Gram-positive bacterial pathogens and commensal Lactobacilli share functional similarity in virulence-associated traits[J].FEMS Immunol Med Microbiol,2007,51(3):526-534.
[26]	LEE J Y,JUNG Y J,JUN H K,et al.Pathogenic potential of Tannerella forsythia enolase[J].Mol Oral Microbiol,2016,31(2):189-203.
[27]	WENG Y D,CHEN F,LIU Y W,et al.Pseudomonas aeruginosa enolase influences bacterial tolerance to oxidative stresses and virulence[J].Front Microbiol,2016,7:1999.

Enolase在鸭疫里默氏杆菌侵袭鸭脑微血管内皮细胞中的作用

Effect of Enolase on Riemerella anatipestifer Invading Duck Brain Microvascular Endothelial Cells

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 27

相关文章 5

编辑推荐

Metrics

本文评价

[1]	李倩倩, 林馨, 韦依琳, 崔昊宇, 邹荣华, 吴晓妮, 葛家振, 黄国梁, 张丽娟, 郑福英, 蔺国珍. RaeR对鸭疫里默氏杆菌外排泵RaeC-RaeA-RaeB的调控作用研究[J]. 畜牧兽医学报, 2023, 54(9): 3793-3802.
[2]	吉果, 陈启伟, 宫晓炜, 刘永生, 郑福英. 鸭疫里默氏杆菌recA基因突变对其生长及DNA损伤反应的影响[J]. 畜牧兽医学报, 2021, 52(9): 2553-2560.
[3]	权衡, 陈启伟, 宫晓炜, 王燕萍, 秦明星, 朱雨佳, 郑福英, 蔺国珍. 鸭疫里默氏杆菌MFS外排泵rant基因缺失株的构建及其介导的耐药性[J]. 畜牧兽医学报, 2021, 52(7): 1991-1999.
[4]	秦明星, 宫晓炜, 陈启伟, 王燕萍, 权衡, 朱雨佳, 郑福英, 蔺国珍. 鸭疫里默氏杆菌RA-LZ01株GE296_RS01450基因介导其对重金属离子产生耐受性的分析[J]. 畜牧兽医学报, 2021, 52(10): 2864-2873.
[5]	姜丹, 冯士彬, 王希春, 李玉, 詹凯, 吴金节, 李锦春. 高温对淮南麻黄鸡血脑屏障的影响[J]. 畜牧兽医学报, 2017, 48(4): 769-776.