【摘要】 通过已建立的兔TLR-2、4mRNA表达水平的SYBR GreenⅠ荧光定量RT-PCR(RRT-PCR)方法,对斯氏艾美尔球虫(E.stiedai)感染后的裂殖生殖期(感染后第8天)、排卵囊初期(感染后第15天)和排卵囊高峰期(感染后第21天),兔肝、脾和外周血单核细胞TLR-2、4mRNA表达水平进行检测。结果显示,TLR-2、4的RRT-PCR产物分别在82.3、79.0℃出现单特异峰,其相关系数分别为0.994 8、0.999 6,扩增效率分别为1.05、0.99。感染E.stiedai后,兔的肝、脾TLR2mRNA表达水平显著高于对照组(P<0.01),且在排卵囊初期最高;TLR4mRNA表达量在裂殖生殖期和排卵囊初期显著下调(P<0.01),而至排卵囊高峰期较对照组显著增加(P<0.01);3个阶段中兔外周血单核细胞TLR2和TLR4mRNA表达水平均显著低于对照组(P<0.01)。本研究提示TLR2和TLR4参与了兔体对E.stiedai的感染应答。更多还原

【Abstract】 In this study,real-time reverse transcription PCR(RRT-PCR)were established to determine the expression of toll like receptor 2(TLR2)and TLR4 in rabbit at 8,15 and 21days post infection(dpi)with Eimeria stiedai.The results showed that the melting temperature of specific products for TLR2 and TLR4 was 82.3and 79.0℃,the amplification efficiency was 105% and99%,and the relative coefficient was 0.994 8and 0.999 6,respectively.The expression of TLR2 mRNA were up-regulated significantly in livers and spleens at 8,15 and 21dpi(P<0.01),and reached the highest level at 15 dpi up to 12.05 and 6.089 folds,respectively.In contrast,the expression levels of TLR4 were down-regulated at the merogony stage(8dpi)and early oocysts shedding stage(15dpi)(P<0.01),however,they were significantly higher than the control at 21 dpi,which is the peak production of oocysts stage.While the expression of TLR2 and TLR4in peripheral mononuclear cells of rabbit infected with E.stiedai were decreased significantly at 8,15 and 21dpi(P<0.01).Taken together,these data indicate that TLR2 and TLR4may play an important role in response to E.stiedai infection.更多还原