rfaE基因通过MAPKs/NF-κB信号通路对副猪嗜血杆菌LOS诱导猪肺泡巨噬细胞炎症反应中作用的研究

doi:10.11843/j.issn.0366-6964.2016.05.015

Abstract

Abstract:

The aim of this study was to study the role of rfaE gene in Haemophilus parasuis lipooligosaccharide (LOS) which induced signal molecules mRNA transcription and MAPKs/ NF-κB signaling pathways in porcine alveolar macrophages (PAMs).The LOS of H.parasuis SC096 strain，rfaE mutant and its complementation was extracted.The PAMs were stimulated with LOS (5 and 10 μg) from H.parasuis SC096，ΔrfaE mutant (ΔrfaE) and its complementation (cΔrfaE) for different time points.The RNA and protein was extracted from the collected cells.The extracted RNA was reversed into cDNA.The mRNA transcription of TLR4，MD2，NF-κB，MAP2K2，ERK，P38 and JNK were then detected by Real-time PCR.The protein of NF-κB p65/ Phospho-NF-κB p65，IκBα，ERK，JNK and p38/ Phospho-p38 were detected by western blot.The results showed that the mRNA transcription of TLR4，MD2，MAP2K2，ERK，P38 and JNK in PAMs induced by 5 and 10 μg HPS-LOS for 6，12 and 24 h were up-regulated，and significantly higher than that in PAMs induced by 5 and 10 μg ΔrfaE-LOS (P<0.05).The mRNA transcription of NF-κB in PAMs induced by 5 and 10 μg HPS-LOS for 6，12 and 24 h were higher than that in PAMs induced by 5 and 10 μg ΔrfaE-LOS，but no significant difference was observed.The protein of IκBα in PAMs induced by 5 and 10 μg HPS-LOS for 6 and 12 h were significantly lower than that in PAMs induced by 5 and 10 μg ΔrfaE-LOS (P<0.05).The p65 phosphorylation，p38 phosphorylation，ERK and JNK in PAMs induced by 5 and 10 μg HPS-LOS for 6 and 12 h were significantly higher than that in PAMs induced by 5 and 10 μg ΔrfaE-LOS (P<0.05).At the same time，the signal molecules mRNA transcription level and the protein level in PAMs induced by cΔrfaE-LOS can restore to the level in PAMs induced by HPS-LOS.The date confirmed that the loss of rfaE gene could effectively reduced the inflammatory process in PAMs induced by HPS-LOS by blocking the MAPKs/ NF-κB signaling pathways.

CLC Number:

S852.613

ZENG Ze，HE Huan，REN Yu-peng，YUE Hua，ZHANG Bin. The Effect of rfaE Gene in Haemophilus parasuis Lipooligosaccharide Induced Inflammation via the MAPKs/ NF-κB Signaling Pathways in Porcine Alveolar Macrophages[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(5): 978-984.

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The Effect of rfaE Gene in Haemophilus parasuis Lipooligosaccharide Induced Inflammation via the MAPKs/ NF-κB Signaling Pathways in Porcine Alveolar Macrophages

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