rfaE基因在副猪嗜血杆菌LOS刺激猪肺泡巨噬细胞炎性因子mRNA转录中的作用

doi:10.11843/j.issn.0366-6964.2015.07.021

畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (7): 1232-1237.doi: 10.11843/j.issn.0366-6964.2015.07.021

rfaE基因在副猪嗜血杆菌LOS刺激猪肺泡巨噬细胞炎性因子mRNA转录中的作用

曾泽，岳华，冯晓辉，何欢，张斌^*

(西南民族大学生命科学与技术学院，成都610041)

收稿日期:2014-10-11 出版日期:2015-07-23 发布日期:2015-07-23
通讯作者: 张斌，副研究员，E-mail: binovy@sina.com
作者简介:曾泽（1991-），女，贵州毕节人，硕士生，主要从事动物病原分子生物学研究，E-mail: 839017291@qq.com
基金资助:
“十二五”国家高技术研究发展（863）计划 (2012AA101304)；国家自然科学基金（31302119）；四川省教育厅项目（14ZB046）；西南民族大学研究生创新型科研项目（CX2014SZ94）；公益性行业（农业）科研专项（201303034-1）

The Effect of rfaE Gene in Haemophilus parasuis LOS Induces Pro-inflammatory Cytokine mRNA Transcription in Porcine Alveolar Macrophages

ZENG Ze，YUE Hua，FENG Xiao-hui，HE Huan，ZHANG Bin^*

(College of Life Science and Technology，Southwest University for Nationalities，Chengdu 610041，China)

Received:2014-10-11 Online:2015-07-23 Published:2015-07-23

摘要/Abstract

摘要：

为研究rfaE基因在副猪嗜血杆菌（HPS）脂寡糖（LOS）刺激猪肺泡巨噬细胞（PAMs）炎性因子（IL-1α、IL-1β、IL-6、IL-8和TNF-α）mRNA转录中的作用，提取HPS SC096株及其rfaE基因缺失株（ΔrfaE）和互补株（cΔrfaE）的LOS，分别用5和10 μg HPS-LOS、ΔrfaE-LOS、cΔrfaE-LOS和E.coli-LPS刺激PAMs，分别在2、4、6、12和24 h后收集细胞，提取RNA并反转录成cDNA，运用RT-PCR检测IL-1α、IL-1β、IL-6、IL-8和TNF-α mRNA的转录水平。结果表明用5 μg HPS-LOS刺激细胞时，2、4、6、12和24 h后IL-1α mRNA的转录水平均升高，显著高于ΔrfaE-LOS刺激细胞后的IL-1α mRNA转录水平（P<0.05），12和24 h后IL-1β、IL-6和IL-8 mRNA的转录水平均升高，显著高于ΔrfaE-LOS刺激细胞后的mRNA转录水平（P<0.05），24 h后TNF-α mRNA的转录水平升高，显著高于ΔrfaE-LOS刺激细胞后的mRNA转录水平（P<0.05）；10 μg HPS-LOS刺激细胞后IL-1α、IL-1β、IL-6、IL-8和TNF-α mRNA的转录水平均升高，显著高于ΔrfaE-LOS刺激细胞后的mRNA转录水平（P<0.05）。同时cΔrfaE-LOS刺激PAMs后IL-1α、IL-1β、IL-6、IL-8和TNF-α mRNA的转录水平能够恢复到HPS-LPS水平。首次证实rfaE基因在HPS LOS刺激PAMs炎性因子mRNA转录水平中起着重要的作用。

Abstract:

The aim of this study was to study the role of rfaE gene of Haemophilus parasuis lipooligosaccharide (LOS) which induced pro-inflammatory cytokine (interleukin (IL)-1α，IL-1β，IL-6，IL-8 and tumor necrosis factor (TNF)-α) mRNA transcription 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 μg and 10 μg) from H.parasuis SC096，ΔrfaE mutant (ΔrfaE) and its complementation (cΔrfaE) and LPS from Escherichia coli for 2，4，6，12 and 24 h.The RNA was extracted and reversed into cDNA.The mRNA transcription of IL-1α，IL-1β，IL-6，IL-8 and TNF-α were then detected by Real-time PCR.The result showed that the mRNA transcription of IL-1α in PAMs induced by 5 μg LOS from H.parasuis for 2，4，6，12 and 24 h were up-regulated，and significantly higher than the mRNA transcription of IL-1α in PAMs induced by 5 μg LOS from ΔrfaE (P<0.05)， the mRNA transcription of IL-1β，IL-6，IL-8 in PAMs induced by 5 μg LOS from H.parasuis for 12 and 24 h were up-regulated，and remarkably higher than the mRNA transcription of IL-1α in PAMs induced by 5 μg LOS from ΔrfaE (P<0.05)，the mRNA transcription of TNF-α in PAMs induced by 5 μg LOS from H.parasuis for 24 h was up-regulated，and notably higher than the mRNA transcription of IL-1α in PAMs induced by 5 μg LOS from ΔrfaE (P<0.05)；the 10 μg LOS from H.parasuis up-regulated mRNA transcription of IL-1α，IL-1β，IL-6，IL-8 and TNF-α in PAMs，and were significantly higher than that of 10 μg LOS from ΔrfaE mutant (P<0.05).At the same time，the cytokine mRNA transcription levels in PAMs induced by LOS from cΔrfaE were restored to the level in PAMs induced by LOS from H.parasuis.The date firstly confirmed that the rfaE gene were involved in the pro-inflammatory cytokine mRNA transcription in PAMs induced by H.parasuis LOS，suggesting that the rfaE gene play an important role in the pathogenesis of H.parasuis LOS.

中图分类号:

S852.613

曾泽，岳华，冯晓辉，何欢，张斌. rfaE基因在副猪嗜血杆菌LOS刺激猪肺泡巨噬细胞炎性因子mRNA转录中的作用[J]. 畜牧兽医学报, 2015, 46(7): 1232-1237.

ZENG Ze，YUE Hua，FENG Xiao-hui，HE Huan，ZHANG Bin. The Effect of rfaE Gene in Haemophilus parasuis LOS Induces Pro-inflammatory Cytokine mRNA Transcription in Porcine Alveolar Macrophages[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(7): 1232-1237.

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rfaE基因在副猪嗜血杆菌LOS刺激猪肺泡巨噬细胞炎性因子mRNA转录中的作用

The Effect of rfaE Gene in Haemophilus parasuis LOS Induces Pro-inflammatory Cytokine mRNA Transcription in Porcine Alveolar Macrophages

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