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  

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

作　　者：何欢[1] 陈新诺 曾泽[1] 任玉鹏[1] 汤承[1] 张斌[1] 岳华[1] 

机构地区：[1]西南民族大学生命科学与技术学院,成都610041

出　　处：《畜牧兽医学报》2016年第7期1428-1434,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31302119);四川省教育厅创新项目(14ZB046);公益性行业(农业)科研专项(201303034-1);西南民族大学研究生创新科研项目(CX2015SZ074)

摘　　要：为了研究副猪嗜血杆菌(Haemophilus parasuis,HPS)弱毒株11型H465株OmpP2刺激猪肺泡巨噬细胞(PAMs)炎性因子mRNA的转录和对NF-κB和MAPKs信号通路的影响,提取并纯化HPS血清11型H465株的OmpP2,分别用5、10μg·mL-1 OmpP2刺激PAMs 6、12h后收集细胞,提取总RNA和蛋白质。将提取的RNA反转录成cDNA,运用RT-PCR检测炎性因子(IL-1α、IL-1β、IL-6、IL-8和TNF-α)mRNA转录水平。利用Western blot方法检测ERK1/2、JNK、P38、P65和IκBα蛋白表达量的变化。结果表明:HPS H465株的OmpP2能够显著地诱导IL-1α、IL-1β、IL-6、IL-8和TNF-α的mRNA转录水平上调(P<0.05),同时能够引起JNK、P65蛋白表达水平显著升高和IκBα蛋白表达水平显著降低(P<0.05)。上述结果证实了HPS血清11型H465株的OmpP2能够通过调节MAPKs信号通路中JNK蛋白以及NF-κB信号通路中P65蛋白和IκBα蛋白降解促进炎性因子IL-1α、IL-1β、IL-6、IL-8和TNF-α的转录。The aim of this study was to explore the role of out membrane protein P2（OmpP2）in Haemophilus parasuis serovar 11 reference H465strain 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 5and 10μg·mL-1 for 6and 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-8and TNF-αwere 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-8and TNF-αin 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 IκBαwas 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 IκBαin NF-κB and MAPKs signaling pathways to promote the pro-inflammatory cytokines expression.

关 键 词：副猪嗜血杆菌 OmpP2 炎性因子 NF-κB和MAPK信号通路 

分 类 号：S852.613[农业科学—基础兽医学]