脂多糖对仔猪外周血白细胞Toll样受体4信号通路关键基因表达的影响  

Effects of Lipopolysaccharide Challenge on the mRNA Expression of Key Genes in TLR4 Signaling Pathway in Peripheral Leucocytes of Weaned Pigs

作　　者：吴欢听 刘玉兰[1] 陈逢[1] 朱惠玲[1] 侯永清[1] 皮定安[1] 冷炜博[1] 

机构地区：[1]武汉轻工大学,动物营养与饲料科学湖北省重点实验室,湖北武汉430023

出　　处：《中国畜牧杂志》2014年第3期60-64,共5页Chinese Journal of Animal Science

基　　金：教育部新世纪优秀人才支持计划(NCET-10-0158);国家自然科学基金面上项目(30972109);湖北省自然科学基金项目(2009CDB006;2010CDA050);湖北省国际科技合作项目(2011BFA008);武汉市科技计划项目(201171034320)

摘　　要：本试验旨在研究免疫应激对仔猪外周血白细胞Toll样受体4(toll-like receptor 4,TLR 4)信号通路关键基因表达水平的影响。选取10头健康的断奶仔猪,分成2个处理,每个处理5个重复。试验组注射100μg/kg体重的脂多糖(LPS)建立免疫应激模型,对照组注射等量生理盐水。注射LPS或生理盐水前(0 h)和注射后1、2、3、4、6、8 h和24 h采血分离白细胞,应用实时荧光定量PCR技术测定TLR4信号通路关键基因,包括TLR4、髓样分化因子88(MyD88)、肿瘤坏死因子受体相关因子6(TRAF6)、核因子-κB(NF-κB)和肿瘤坏死因子-α(TNF-α)在白细胞中mRNA的表达水平。结果表明:TLR4在LPS注射后2、4 h和6 h表达水平显著降低(P﹤0.05);MyD88在LPS注射后1、3 h和8 h显著降低(P﹤0.05);TRAF6及NF-κB在LPS注射后3 h显著降低(P﹤0.05);TNF-α在LPS注射后6、8和24 h显著升高(P﹤0.05)。这表明LPS注射后短时间内,白细胞对LPS产生早期暂时耐受性,从而阻碍炎症反应的发生。This experiment was conducted to investigate the effect of immunological challenge on the mRNA expression pattern of the key genes in toll-like receptor 4 （TLR4） signaling pathway in peripheral leucocytes of weaned pigs. Ten Pigs were allotted to two groups with 5 replicates per treatment and 1 pig per replicate. The pigs in the treatment group were injected with 100 p,g/kg BW lipopolysaccharide （LPS）, whereas pigs in the control group were injected with an equivalent amount of sterile saline. Blood samples were collected to isolate leucocytes before LPS challenge （0 h）, and at 1, 2, 4, 6, 8 and 24 h after LPS challenge. The mRNA expression levels of the key genes in TLR4 signaling pathway, including TLR4, myeloid differentiation factor 88 （MyD88）, TNF-α receptor-associated factor 6 （TRAF6）, nuclear factor-KB （NF-KB） and tumor necrosis factor-or （TNF-α） in peripheral leucocytes, were detected by Real-time quantitative PCR. TLR4 mRNA expression level was reduced significantly at 2, 4 and 6 h after LPS challenge （P〈0.05）. MyD88 mRNA expression level was decreased significantly at 1, 3 and 8 h after LPS challenge （P〈0.05）. TRAF6 and NF-KB mRNA expression level were decreased significantly at 3 h after LPS challenge （P〈0.05）. TNF-α mRNA expression level was increased significantly at 6, 8 and 24 h after LPS challenge （P〈0.05）. These results indicate that the leucocytes show an early temporary tolerance to LPS, hampering the inflammatory response at a short time after LPS challenge.

关 键 词：脂多糖 外周血白细胞 TLR4信号通路 仔猪 

分 类 号：S828.5[农业科学—畜牧学]