副猪嗜血杆菌OmpP2刺激猪肺泡巨噬细胞炎性因子mRNA转录及致炎机制初步分析

doi:10.11843/j.issn.0366-6964.2016.07.016

Abstract

Abstract:

The aim of this study was to explore the role of out membrane protein P2 (OmpP2) in Haemophilus parasuis serovar 11 reference H465 strain to induce pro-inflammatory cytokine mRNA transcription，and NF-κB and MAPKs signaling pathways in porcine alveolar macrophages (PAMs).The OmpP2 was extracted from the H.parasuis H465 and stimulated in PAMs with 5 and 10 μg•mL－1 for 6 and 12 hours.The total RNA and cells protein were extracted and the RNA was reversed into cDNA.The mRNA transcription levels of IL-1α，IL-β，IL-6，IL-8 and TNF-α were detected by Real-time PCR，and the protein of NF-κB P65，IκBα，ERK，JNK and P38 were detected by Western blot.The result showed that the mRNA transcription of IL-1α，IL-β，IL-6，IL-8 and TNF-α in PAMs induced by the OmpP2 from H.parasuis were significantly up-regulated (P<0.05)，and the protein of JNK，P65 in PAMs induced by the OmpP2 from H.parasuisis significantly increased and the protein of IκBα was significantly declined(P<0.05).The above results exhibit that the OmpP2 could effectively activate the JNK and P65 proteins and increase the degradation of IκBα in NF-κB and MAPKs signaling pathways to promote the pro-inflammatory cytokines expression.

CLC Number:

S852.613

HE Huan，CHEN Xin-nuo，ZENG Ze，REN Yu-peng，TANG Cheng，ZHANG Bin，YUE Hua. The Initial Research of OmpP2 in Haemophilus parasuis Induces Pro-inflammatory Cytokine mRNA Transcription and Inflammatory Mechanism in Porcine Alveolar Macrophages[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(7): 1428-1434.

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References

［1］CAI X，CHEN H，BLACKALL P J，et al.Serological characterization of Haemophilus parasuis isolates from China［J］.Vet Microbiol，2005，111(3-4)：231-236.
［2］RAPP-GABRIELSON V J，GABRIELSON D A.Prevalence of Haemophilus parasuis serovars among isolates from swine［J］. Am J Vet Res，1992，53(5)：659-664.
［3］PAGÈS J M，JAMES C E，WINTERHALTER M.The porin and the permeating antibiotic：a selective diffusion barrier in Gram-negative bacteria［J］.Nat Rev Microbiol，2008，6(12)：893-903.
［4］赵倩，汤承，杨发龙，等.副猪嗜血杆菌ompP2基因的结构特征及其与毒力的联系［J］.中国科学：生命科学，2010，40(6)：522-532.
ZHAO Q，TANG C，YANG F L，et al.The correlation between structural characterization of ompP2 gene and virulence of Haemophilus parasuis［J］.Scientia Sinica：Vitae，2010，40(6)：522-532.(in Chinese)
［5］ZHANG B，XU C，ZHANG L，et al.Enhanced adherence to and invasion of PUVEC and PK-15 cells due to the overexpression of RfaD，ThyA and Mip in the ΔompP2 mutant of Haemophilus parasuis SC096 strain［J］.Vet Microbiol，2013，162(2-4)：713-723.
［6］ZHOU S，HE X，XU C，et al.The outer membrane protein P2 (OmpP2) of Haemophilus parasuis induces proinflammatory cytokine mRNA expression in porcine alveolar macrophages［J］.Vet J，2014，199(3)：461-464.
［7］CINAR M U，ISLAM M A，UDDIN M J，et al.Evaluation of suitable reference genes for gene expression studies in porcine alveolar macrophages in response to LPS and LTA［J］.BMC Res Notes，2012，5：107.
［8］LIVAK K，SCHMITTGEN T D.Analysis of relative gene expression date using real-time quantitative PCR and the 2-ΔΔCT method［J］.Methods，2001，25(4)：402-408.
［9］RUIZ A，OLIVEIRA S，TORREMORELL M，et al.Outer membrane proteins and DNA profiles in strains of Haemophilus parasuis recovered from systemic and respiratory sites［J］.J Clin Microbiol，2001，39(5)：1757-1762.
［10］陆承平.兽医微生物学［M］.北京：中国农业出版社，2007：144-146.
LU C P.Veterinary microbiology［M］.Beijing：China Agriculture Press，2007：144-146.(in Chinese)
［11］OLIVEIRA S，PIJOAN C.Haemophilus parasuis：new trends on diagnosis，epidemiology and control［J］.Vet Microbiol，2004，99(1)：1-12.
［12］VAHLE J L，HAYNES J S，ANDREWS J J.Experimental reproduction of Haemophilus parasuis infection in swine：clinical，bacteriological，and morphologic findings［J］.J Vet Diagn Invest，1995，7(4)：476-480.
［13］GALDIERO M，D’AMICO M，GORGA F，et al. Haemophilus influenzae porin contributes to signaling of the inflammatory cascade in rat brain［J］.Infect Immun，2001，69(1)：221-227.
［14］RAY A，CHATTERJEE N S，BHATTACHARYA S K，et al.Porin of Shigella dysenteriae enhances mRNA levels for Toll-like receptor 2 and MyD88，up-regulates CD80 of murine macrophage，and induces the release of interleukin-12［J］.FEMS Immunol Med Microbiol，2003，39(3)：213-219.
［15］MASSARI P，GUNAWARDANA J，LIU X，et al.Meningococcal porin PorB prevents cellular apoptosis in a toll-like receptor 2- and NF-kappaB-independent manner［J］.Infect Immun，2010，78(3)：994-1003.
［16］KIM H J，LEE H S，CHONG Y H，et al.p38 mitogen-activated protein kinas up-regulates LPS-induced NF-κB activation in the development of lung injury and RAW 264.7 macrophages［J］.Toxicology，2006，225(1)：36-47.
［17］RAVANTI L，TORISEVA M，PENTTINEN R，et al.Expression of human collagenase-3(MMP-13)by fetal skin fibroblasts is induced by transforming growth factor β via p38 mitogen-activated protein kinase［J］.FASEB J，2001，15(6)：1098-1100.
［18］LIU Y W，CHEN C C，TSENG H P，et al.Lipopolysaccharide-induced transcriptional activation of interleukin-10 is mediated by MAPK- and NF-kappaB-induced CCAAT /enhancer-binding protein delta in mouse macrophages［J］. Cell Signal，2006，18(9)：1492- 1500.
［19］LEE J I，BURCKART G J.Nuclear factor kappa B：important transcription factor and therapeutic target［J］.J Clin Pharmacol，1998，38(11)：981-993.
［20］CHANG L，KARIN M.Mammalian MAP kinase signalling cascades［J］.Nature，2001，410(6824)：37-40.

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The Initial Research of OmpP2 in Haemophilus parasuis Induces Pro-inflammatory Cytokine mRNA Transcription and Inflammatory Mechanism in Porcine Alveolar Macrophages

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