细菌内毒素对鸡NLRC5受体及IFN基因转录的影响  

Transcription Change of Chicken NLRC5 Receptor and IFN Genes Initiated by Bacterial Endotoxins

作　　者：常国斌[1] 陈蓉[1] 栾德琴[1] 张颖[1] 刘向萍[2] 马腾[1] 戴爱琴[1] 周伟[1] 陈国宏[1] 

机构地区：[1]扬州大学动物科学与技术学院,扬州225009 [2]中国农业科学院家禽研究所,扬州225003

出　　处：《畜牧兽医学报》2011年第11期1598-1604,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(30972088;31172199);江苏省自然科学基金(BK2009190);江苏省农业科技支撑计划(BE2010375)

摘　　要：本研究旨在探讨鸡NLRC5受体及IFN基因对不同细菌内毒素的诱导表达变化,为禽类NLRC5受体的分子功能及其参与先天免疫调控的作用机制提供理论依据。用不同浓度的(0、0.1、1.0、10.0μg.mL-1)来自致病性大肠杆菌(E.coli,EC)和肠炎沙门氏菌(S.enteriditis,SE)的2种内毒素(LPS)分别刺激鸡天然巨噬细胞系(HD11)2、4和6h,在不同的时间点收集细胞,提取总RNA,并设计相关引物,利用荧光定量PCR分析鸡NLRC5受体以及下游功能基因IFNA和IFNB的转录量变化,探讨HD11细胞对不同来源的细菌内毒素的免疫反应。研究表明:与对照组相比,1.0和10.0μg.mL-1的EC-LPS、SE-LPS均能诱导HD11细胞产生免疫应答,但0.1μg.mL-1组未能诱导成功;不同细菌LPS诱导的免疫应答时间不一致,经EC-LPS诱导的NLRC5和IFNB基因均在刺激后6h达到转录高峰,与2、4h差异显著(P<0.05),与对照组差异极显著(P<0.01);但IFNA基因在2、6h达到高峰,而在4h下降,呈"V"型转录趋势。SE-LPS诱导的NLRC5和IFNA、IFNB转录量随时间均保持一直上升的趋势,在6h达到高峰,且与对照组及2、4h差异显著(P<0.05)。结果表明NLRC5受体与IFNA和IFNB共同参与了细胞对细菌内毒素的免疫应答,NLRC5基因可能对鸡先天免疫功能具有调节作用。This study was conducted to investigate the transcription change of chicken NLRC5 receptor and IFN genes initiated by different bacterial endotoxin,aiming to provide a theoretical basis on poultry NLRC5 receptor function and its regulation mechanism on innate immune.Different concentrations（0,0.1,1.0,10.0 μg·mL-1） of endotoxin from Escherichia coli（E.coli,EC） and Salmonella enteritidis（S.enteriditis,SE） were used to stimulate the chicken macrophage cell line（HD11） for 2,4 and 6 hours,respectively.Total RNA was isolated from cells at different time points,then the transcription difference of NLRC5,IFNA and IFNB genes between different treatments were determined by quantitative real-time RT-PCR,using SYBR Green RT-PCR,and immune response for HD11 cells to different bacterial endotoxin were finally analyzed.In comparison with the control group,1.0 and 10.0 μg·mL-1 of the EC-LPS,SE-LPS could induce immune responses in HD11 cells,whereas 0.1 μg·mL-1 group failed to induce responses successfully.The immune response time induced by different bacterial LPS was different.NLRC5 and IFNB gene transcription induced by EC-LPS,SE-LPS showed up-regulation and reached the peak at 6 hours post-stimulation,which presented significant difference with that of 2,4 hours post-stimulation,but IFNA gene transcription induced by EC-LPS reached the peak at 2,6 hours,while was down regulated at 4 hours post-stimulation,showing as ＂V＂-type transcription trend.The transcription of all three genes induced by SE-LPS reached the peak at 6 hours post-stimulation and presented significant difference with that of 2,4 hours and control group,showing as one up trend line with time going on.The results showed that the IFNA,IFNB and NLRC5 receptors participated jointly the immune response to bacterial endotoxins,and NLRC5 genes may have regulate function on innate immune in chicken.

关 键 词：NLRC5 IFN 鸡 先天免疫 调节 

分 类 号：S852.4[农业科学—基础兽医学]