运输应激对猪淋巴结IL-2、IL-6和IL-10及其受体mRNA转录的影响  

Effects of transportation stress on transcription of IL-2,IL-6 and IL-10 and the relative receptors mRNA of lymph nodes in pig

作　　者：吕琼霞[1] 张书霞[1] 赵茹茜[1] 

机构地区：[1]南京农业大学动物医学院/农业部动物生理生化重点开放实验室,江苏南京210095

出　　处：《南京农业大学学报》2011年第1期95-100,共6页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金重点项目(30430420)

摘　　要：为研究运输应激对猪肠系膜淋巴结中IL-2、IL-6、IL-10及其受体mRNA转录的影响,并探讨p38丝裂原活化蛋白激酶(P38MAPK)和核因子κB(NF-κB)对其转录的调控作用,用荧光定量RT-PCR方法对肠系膜淋巴结中3种细胞因子及其受体mRNA进行定量检测,用ELISA方法检测猪肠系膜淋巴结组织匀浆液中P38MAPK含量的动态变化,用激光扫描共聚焦显微镜观察运输前后NF-κB在肠系膜淋巴结细胞中的分布。结果显示:经1、2和4 h的运输应激,肠系膜淋巴结中3种细胞因子的转录均发生了显著变化,且均是4 h时达到最大值,并极显著高于对照组、1 h组和2 h组(P<0.01),而其相应受体的变化趋势不同。NF-κB在肠系膜淋巴结细胞中的表达呈逐渐上升趋势,到4 h时达到最大,其入核率也是同样的变化趋势。运输应激过程中P38MAPK含量没有明显的变化。结论:运输应激可引起猪肠系膜淋巴结细胞因子及其受体mRNA转录的急剧变化,且存在时效性;NF-κB的激活是调控猪肠系膜淋巴结相关细胞因子变化的一个重要因素。The objective of this study was to explore the transcription of IL-2,IL-6,IL-10 and relative receptors mRNA in mesenteric lymph nodes and regulation of P38MAPK and NF-κB in pigs during transportation stress.SYBR GreenⅠbased one-step real-time quantitative RT-PCR amplification was performed in quantitation of the three cytokines and their receptors mRNAs.P38MAPK in the homogenization sample of mesenteric lymph nodes was measured with homologous specific ELISA.The distribution of NF-κB in the mesenteric lymph nodes was examined using confocal laser scanning microscope（CLSM）.After 1,2 and 4 h of transportation,the expression level of three cytokines and relative receptors mRNAs in mesenteric lymph nodes was significantly changed compared with the control group and got to maximum at 4 h during transportation stress,and significantly higher than the control,1 h and 2 h groups（P0.01）,but the trend of changes in their corresponding receptors are different.The expression level of NF-κB in mesenteric lymph node cells showed a gradual upward trend and rose to maximum at 4 h and the nucleation rate was the same trend,but there were no significant changes with respect to P38MAPK during the transport stress.The cytokines and receptors mRNA could be altered by transportation stress and was also related to the timing of post-stress in pigs.The combination of transcription and regulation of cytokines signaling mechanism showed that the activation of NF-κB in the mesenteric lymph nodes was an important factor in the peripheral immune-related cytokines change in transportation stress pigs.

关 键 词：猪 运输应激 肠系膜淋巴结 细胞因子 P38MAPK 核因子ΚB 

分 类 号：S852.4[农业科学—基础兽医学]