TLRs介导的信号通路在马链球菌感染RAW264.7细胞中的作用  

Effect of TLRs Mediated Signal Pathway on Streptococcus equi Infection in RAW264.7 Cells

作　　者：杜润慈[1] 刘建华[1] 史茜[2] 付强[1] 冉多良[1] 

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052 [2]新疆出入境检验检疫局动植物检疫中心,乌鲁木齐830063

出　　处：《中国畜牧兽医》2016年第5期1270-1276,共7页China Animal Husbandry & Veterinary Medicine

基　　金：国家科技支撑计划项目(2012BAD46B00)

摘　　要：为探究Toll样受体(TLRs)介导的信号通路在马链球菌马亚种(S.equi)感染小鼠巨噬细胞RAW264.7中的作用,收集S.equi感染后不同时间点的RAW264.7细胞,提取总RNA并反转录成cDNA,利用实时荧光定量PCR技术检测细胞Toll样受体1、2、6(TLR1、TLR2、TLR6)、接头蛋白骨髓分化蛋白88(MyD88)及细胞因子IL-1、IL-6、IL-10、IL-12、TNF-αmRNA的表达情况。结果显示,S.equi感染RAW264.7细胞后6h时,TLR1、TLR2、TLR6与MyD88mRNA水平均较对照组没有显著差异(P>0.05);感染后12h时,TLR1、TLR2和TLR6mRNA表达量未出现明显上升(P>0.05),而MyD88mRNA水平极显著升高(P<0.01);感染后24h时,TLR1、TLR2和TLR6mRNA表达水平出现极显著升高(P<0.01),MyD88 mRNA表达没有显著变化(P>0.05),且IL-10和IL-12mRNA水平与对照组相比极显著升高(P<0.01),IL-1、IL-6和TNF-αmRNA水平均极显著下降(P<0.01)。结果表明,TLRs介导的信号通路参与S.equi感染RAW264.7细胞的免疫应答反应。To investigate the effects of Toll-like receptors （TLRs） mediated signaling pathway in mouse macrophage RAW264.7 cells on Streptococcus equi subspecies equi （S.equi） infection,RAW264.7 cells were infected with S.equi for different time intervals.The mRNA expression levels of TLR1,TLR2,TLR6,myeloid differentiation factor 88 （MyD88）,IL-1,IL-6,IL-10,IL-12 and TNF-α were detected by fluorescence quantitative Real-time PCR.The results showed that at 6 h post-infection,the expression levels of TLR1,TLR2,TLR6 and MyD88 in S.equi infected RAW264.7 cells had no significant difference compared with control group （P＞0.05）.At 12 h post-infection,the mRNA expression levels of TLR1,TLR2 and TLR6 had no obviously change （P＞0.05）,while MyD88 mRNA expression level was extremely significantly increased （P＜ 0.01）.At 24 h post-infection,the mRNA levels of TLR1,TLR2,TLR6 were extremely significantly increased （P＜0.01）,the mRNA level of MyD88 had no significant difference （P＞0.05）,meanwhile,compared with uninfected cells,IL-10 and IL-12 mRNA expression were extremely significant increased in S.equi infected RAW264.7 cells （P＜0.01）,but IL-1,IL-6 and TNF-α mRNA levels were extremely significantly decreased （P＜0.01）.These results indicated that the signaling pathway mediated by TLRs participated in the immune response against S.equi.

关 键 词：马链球菌马亚种 RAW264.7细胞 TOLL样受体 骨髓分化蛋白88 细胞因子 

分 类 号：Q2-33[生物学—细胞生物学]