牛A型巴氏杆菌高、低毒力株LPS激活RAW264.7细胞TLR4信号通路的比较分析

doi:10.11843/j.issn.0366-6964.2019.03.017

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (3): 620-626.doi: 10.11843/j.issn.0366-6964.2019.03.017

牛A型巴氏杆菌高、低毒力株LPS激活RAW264.7细胞TLR4信号通路的比较分析

李艳红, 刘洁, 李伟平, 崔雨婷, 彭远义, 吴正理*

西南大学动物科技学院, 重庆 400715

收稿日期:2018-08-20 出版日期:2019-03-23 发布日期:2019-03-23
通讯作者: 吴正理,主要从事病原微生物与免疫研究,E-mail:zh20140202@swu.edu.cn
作者简介:李艳红(1980-),女,讲师,博士,主要从事病原微生物与免疫研究,Tel:023-68251070,E-mail:lyhong08@126.com
基金资助:
国家自然科学基金（31770965；31600716）；重庆市基础科学与前沿技术研究专项（cstc2017jcyjAX0420）；中央高校基本科研业务费专项资金（SWU114011；SWU114015）；国家现代农业（肉牛牦牛）产业技术体系建设专项基金（CARS-37）

Comparative Analysis of TLR4 Signaling Pathway in RAW264.7 Cells Activated by LPS from High-and Low-virulence Strains of Bovine Pasteurella multocida Serotype A

LI Yanhong, LIU Jie, LI Weiping, CUI Yuting, PENG Yuanyi, WU Zhengli*

College of Animal Science and Technology, Southwest University, Chongqing 400715, China

Received:2018-08-20 Online:2019-03-23 Published:2019-03-23

摘要/Abstract

摘要：

本研究旨在比较牛A型多杀性巴氏杆菌高毒力株（PmCQ2）和低毒力株（PmCQ6）脂多糖（LPS）对小鼠巨噬细胞RAW264.7内TLR4信号通路的影响。选用体外培养RAW264.7细胞，经LPS刺激后检测细胞内TLR4信号激活及对TNF-α和IL-12p40表达的影响；并利用Western blot检测LPS对IκBα磷酸化及NF-κBp65活化的影响。结果显示，高毒力株LPS_PmCQ2和低毒力株LPS_PmCQ6分别刺激RAW264.7细胞后，均能显著提高TLR4的表达，并诱导细胞因子TNF-α和IL-12p40的分泌表达；LPS_PmCQ2和LPS_PmCQ6均能诱导IκBα磷酸化，促进NF-κBp65核转位，但二者没有显著差异（P>0.05）。本研究表明牛A型多杀性巴氏杆菌高、低毒力株LPS均能参与TLR4介导的小鼠巨噬细胞免疫应答，且二者对TLR4介导的IκBα-NF-κB信号通路的作用无显著差异，暗示牛A型多杀性巴氏杆菌对小鼠巨噬细胞的毒力与LPS无明显相关性，可能由其他毒力因子决定。

Abstract:

This study was designed to investigate the effects of TLR4 signaling pathway in RAW264.7 cells treated with lipopolysaccharide (LPS) in two different virulent Pasteurella multocida serotype A isolates (high-virulent PmCQ2 and low-virulent PmCQ6) respectively. The activation of TLR4 pathway, expression of TNF-α and IL-12p40, phosphorylation of IκBα, and nuclear translocation of NF-κB, were identified in LPS-stimulated RAW264.7 cells. Results showed that the expression of TLR4, TNF-α and IL-12p40 was significantly increased in LPS-stimulated RAW264.7 cells. Furthermore, both LPS_PmCQ2 and LPS_PmCQ6 up-regulated IκBα phosphorylation, and activated TLR4-triggered nuclear translocation of NF-κBp65, but there was no significant difference between the two treatments. The LPS of bovine Pasteurella multocida Serotype A high-and low-virulent strains can induce the TLR4-mediated immune response, and there is no difference in TLR4-mediated IκBα-NF-κB signaling pathway, indicating that there may be no significant difference in virulence between LPS_PmCQ2 and LPS_PmCQ6 on RAW264.7 cells. These results suggested that for RAW264.7 cells, the virulence of Pasteuralla multocida is dependent on others virulence factors, not LPS.

中图分类号:

S852.612

李艳红, 刘洁, 李伟平, 崔雨婷, 彭远义, 吴正理. 牛A型巴氏杆菌高、低毒力株LPS激活RAW264.7细胞TLR4信号通路的比较分析[J]. 畜牧兽医学报, 2019, 50(3): 620-626.

LI Yanhong, LIU Jie, LI Weiping, CUI Yuting, PENG Yuanyi, WU Zhengli. Comparative Analysis of TLR4 Signaling Pathway in RAW264.7 Cells Activated by LPS from High-and Low-virulence Strains of Bovine Pasteurella multocida Serotype A[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(3): 620-626.

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参考文献

[1] WILSON B A, HO M F. Pasteurella multocida:from zoonosis to cellular microbiology[J]. Clin Microbiol Rev, 2013, 26(3):631-655.
[2] DABO S M, TAYLOR J D, CONFER A W. Pasteurella multocida and bovine respiratory disease[J]. Anim Health Res Rev, 2007, 8(2):129-150.
[3] HATFALUDI T, AL-HASANI K, BOYCE J D, et al. Outer membrane proteins of Pasteurella multocida[J]. Vet Microbiol, 2010, 144(1-2):1-17.
[4] 李能章, 刘舒萍,程蓉,等. 牛A型多杀性巴氏杆菌新型保护性抗原的筛选及鉴定[J]. 中国兽医科学, 2016, 46(11):1438-1443.
LI N Z, LIU S P, CHENG R, et al. Screening and identification of new protective antigens of bovine Pasteurella multocida serotype A[J]. Chinese Veterinary Science, 2016, 46(11):1438-1443. (in Chinese)
[5] BOYCE J D, SEEMANN T, ADLER B, et al. Pathogenomics of Pasteurella multocida[M]//AKTORIES K, ORTH J H C, ADLER B. Pasteurella Multocida:Molecular Biology, Toxins and Infection. Berlin, Heidelberg:Springer, 2012, 361:23-38.
[6] EWERS C, LVBKE-BECKER A, BETHE A, et al. Virulence genotype of Pasteurella multocida strains isolated from different hosts with various disease status[J]. Vet Microbiol, 2006, 114(3-4):304-317.
[7] HARPER M, BOYCE J D, ADLER B. Pasteurella multocida pathogenesis:125 years after Pasteur[J]. FEMS Microbiol Lett, 2006, 265(1):1-10.
[8] KATOCH S, SHARMA M, PATIL R D, et al. In vitro and in vivo pathogenicity studies of Pasteurella multocida strains harbouring different ompA[J]. Vet Res Commun, 2014, 38(3):183-191.
[9] DU H H, FANG R D, PAN T T, et al. Comparative genomics analysis of two different virulent bovine Pasteurella multocida isolates[J]. Int J Genomics, 2016, 2016:4512493.
[10] MAY B J, ZHANG Q, LI L L, et al. Complete genomic sequence of Pasteurella multocida, Pm70[J]. Proc Natl Acad Sci U S A, 2001, 98(6):3460-3465.
[11] RAETZ C R H, WHITFIELD C. Lipopolysaccharide endotoxins[J]. Annu Rev Biochem, 2002, 71:635-700.
[12] HILDEBRAND D, SAHR A, WÖLFLE S J, et al. Regulation of Toll-like receptor 4-mediated immune responses through Pasteurella multocida toxin-induced G protein signalling[J]. Cell Commun Signal, 2012, 10(1):22.
[13] KUBATZKY K F, KLOOS B, HILDEBRAND D. Signaling cascades of Pasteurella multocida toxin in immune evasion[J]. Toxins, 2013, 5(9):1664-1681.
[14] GALDIERO M, FOLGORE A, NUZZO I, et al. Neutrophil adhesion and transmigration through bovine endothelial cells in vitro by protein H and LPS of Pasteurella multocida[J]. Immunobiology, 2000, 202(3):226-238.
[15] TAPPING R I, AKASHI S, MIYAKE K, et al. Toll-like receptor 4, but not toll-like receptor 2, is a signaling receptor for Escherichia and Salmonella lipopolysaccharides[J]. J Immunol, 2000, 165(10):5780-5787.
[16] KUBATZKY K F. Pasteurella multocida and immune cells[M]//AKTORIES K, ORTH J H C, ADLER B. Pasteurella Multocida:Molecular Biology, Toxins and Infection. Berlin, Heidelberg:Springer, 2012, 361:53-72.
[17] 杨宝凤, 李能章,邹灵秀,等. 6株牛源A型多杀性巴氏杆菌的分离与鉴定[J]. 中国预防兽医学报, 2014, 36(6):487-489.
YANG B F, LI N Z, ZOU L X, et al. Identification and partial biological characteristics of six serotype A Pasteurella multocida from beef cattle[J]. Chinese Journal of Preventive Veterinary Medicine, 2014, 36(6):487-489. (in Chinese)
[18] IOVANE G, PAGNINI P, GALDIERO M, et al. Role of Pasteurella multocida porin on cytokine expression and release by murine splenocytes[J]. Vet Immunol Immunopathol, 1998, 66(3-4):391-404.
[19] MACNEIL A J, JIAO S C, MCEACHERN L A, et al. MAPK kinase 3 is a tumor suppressor with reduced copy number in breast cancer[J]. Cancer Res, 2014, 74(1):162-172.
[20] WU Z L, LI Y J, MACNEIL A J, et al. Calcineurin-Rcan1 interaction contributes to stem cell factor-mediated mast cell activation[J]. J Immunol, 2013, 191(12):5885-5894.
[21] 华瑞其,赵新新,程安春. 多杀性巴氏杆菌脂多糖的结构与功能研究进展[J]. 畜牧兽医学报, 2016, 47(10):1961-1968.
HUA R Q,ZHAO X X,CHENG A C. Research progress in the lipopolysaccharide of Pasteurella multocida[J]. Acta Veterinaria et Zootechnica Sinica, 2016, 47(10):1961-1968.(in Chinese)
[22] HARPER M, BOYCE J D, COX A D, et al. Pasteurella multocida expresses two lipopolysaccharide glycoforms simultaneously, but only a single form is required for virulence:Identification of two acceptor-specific heptosyl I transferases[J]. Infect Immun, 2007, 75(8):3885-3893.
[23] PRIYA G B, NAGALEEKAR V K, MILTON A A P, et al. Genome wide host gene expression analysis in mice experimentally infected with Pasteurella multocida[J]. PLoS One, 2017, 12(7):e0179420.
[24] SKAUG B, JIANG X M, CHEN Z J. The role of ubiquitin in NF-κB regulatory pathways[J]. Annu Rev Biochem, 2009, 78:769-796.

牛A型巴氏杆菌高、低毒力株LPS激活RAW264.7细胞TLR4信号通路的比较分析

Comparative Analysis of TLR4 Signaling Pathway in RAW264.7 Cells Activated by LPS from High-and Low-virulence Strains of Bovine Pasteurella multocida Serotype A

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