感染Mo对贵州不同品种羊TLRs TRIF通路因子基因转录水平及IFN含量的影响

Effects of Mo infection on transcription levels of TLRs TRIF pathway factor genes and IFN content in sheep of different Guizhou breeds

为了探讨感染绵羊肺炎支原体(Mo)对贵州不同品种羊Toll样受体(TLRs)结构域衔接蛋白(TRIF)信号通路因子TRIF、TRAF6、IRF3、IRF7基因转录水平及干扰素(IFN)含量的影响,试验选择贵州地区的5个主要品种羊,每个品种随机分为对照组(2只)和感染组(3只),感染组试验羊人工感染Mo,用PCR方法检测不同品种羊感染Mo的情况,采用TaqMan RT-qPCR方法检测不同品种羊血液样本中TLRs TRIF信号通路因子基因转录水平,并用ELISA方法检测不同品种羊血液样本中的IFN含量。结果表明:从各品种羊鼻拭子中均检测出与预期大小相符的特异性条带。与对照组相比,相同品种羊在感染Mo后血液样本中TLRs TRIF信号通路因子基因转录水平均出现不同程度的上升,其中以TRIF、IRF7基因转录水平上调较明显(P<0.05或P<0.01);感染后96小时时贵州白山羊和贵州黑山羊TRIF基因转录水平极显著高于其余3个品种羊(P<0.01);感染后96小时时贵州黑山羊和波尔山羊TRAF6基因转录水平显著高于其余3个品种羊(P<0.05);感染后48,96小时时贵州白山羊、黔北麻羊IRF3基因转录水平极显著高于其余3个品种羊(P<0.01);感染后48,96小时时贵州白山羊和贵州黑山羊RIF7基因转录水平均显著高于波尔山羊和湖羊(P<0.05),极显著高于黔北麻羊(P<0.01)。贵州白山羊、黔北麻羊、湖羊感染Mo后随着时间的延长血液中IFN-β含量大体上呈上升趋势,其中黔北麻羊上升幅度最大。说明不同品种羊感染Mo后血液样本中TLRs TRIF信号通路因子基因转录水平及IFN含量均存在差异,出现这种差异的原因可能是Mo对不同品种羊的致病机理不同。

To investigate the effects of Mycoplasma ovipneumoniae(Mo)infection on the transcription levels of TRIF,TRAF6,IRF3 and IRF7 genes of Toll-like receptor(TLRs)domain junction protein(TRIF)signaling pathway and interferon(IFN)content in sheep of different guizhou breeds.The experiment was chosen in five major varieties of sheep in Guizhou,each of which was randomly divided into the control group(two sheep)and the infection group(three sheep).The infection group was artificially infected with Mo.PCR method was used to detect the infection of Mo in different breeds of sheep.TaqMan RT-qPCR method was used to detect the transcription levels of TLRs TRIF signaling pathway factors in blood samples of sheep of different breeds,and ELISA method was used to detect the IFN content in blood samples of sheep of different breeds.The results showed that the specific bands that were consistent with the expected size were detected from each variety of sheep’s nasal swab.Compared with the control group,the transcription levels of TLRs TRIF signal pathway factor genes in blood samples of the same breed of sheep after Mo infection were increased to varying degrees,among which the transcription levels of TRIF and IRF7 genes were significantly up-regulated(P<0.05 or P<0.01).At 96 h after infection,the multiple difference of TRIF gene transcription between Guizhou white goats and Guizhou black goats was significantly higher than that of the other three breeds(P<0.01).At 96 h after infection,the multiple difference of TRAF6 gene transcription between Guizhou black goats and Boer goats was significantly higher than that of the other three breeds(P<0.05).At 48 and 96 hours after infection,the multiple difference of IRF3 gene transcription of Guizhou white goats and Qianbei Pockmarked goats was significantly higher than that of the other three breeds(P<0.01).At 48 and 96 hours after infection,the differences of RIF7 gene transcription levels in Guizhou white and Guizhou black goats were significantly higher than those in Boer and Hu goats(P<0.05),and extremely significantly higher than those in Qianbei Pockmarked goats(P<0.01).After Mo infection,the serum IFN-βcontent of Guizhou white goats,Guizhou Qianbei Pockmarked goats and Hu sheep generally increased with the prolongation of time,and the increase of Guizhou Qianbei Pockmarked goats was the largest.It indicated that there were differences in gene transcription levels of TLRs TRIF signaling pathway factors and IFN content in blood samples of sheep of different breeds infected with Mo,which might be caused by different pathogenic mechanisms of Mo on different breeds of sheep.