中文摘要：

为确定toll样受体家族在绵羊肺泡巨噬细胞的分布及脂多糖（LPS）刺激过程中TLR2、4的动态表达规律。通过RT-PCR方法检测绵羊TLR1-TLR10表达分布;以α-tubulin和GAPDH为内标基因,对绵羊TLR2、4进行克隆和测序,将重组质粒作为标准品建立实时定量PCR标准曲线,通过实时定量PCR方法检测LPS诱导0-48 hTLR2、4的动态表达。结果表明,在绵羊肺泡巨噬细胞中检测到TLR1、2、4、6、7、8、9、10的表达,而TLR3、5未见表达;α-tubulin和GAPDH在LPS诱导前后表达量存在显著差异,均不适合作为内标基因,以使用的细胞数和总RNA量进行均一化校正,发现TLR2、4在诱导后20 min就达到高峰,且在整个诱导过程中维持较高水平。绵羊肺泡巨噬细胞TLR2、4对LPS刺激的应答反应很灵敏,参与机体的早期应答反应。

英文摘要：

To determine the distribution of toll-like receptor family （TLRs） in sheep alveolar macrophage （AM） and the expression pattern of TLR2,4 during lipopolysaccharide （LPS） stimulation. Reverse transcription PCR （ RT- PER） Was used to detect the distribution of sheep TLR1 - TLRIO gene expression, α-tubulin and GAPDH were testified as valid house keeping genes（HKG）. Real -time PCR standard curve was established with sheep TLR2,4 genes recombinant plasmid as a standard criteria,dynamic expression of sheep TLR2,4 in AM with LPS-induced 0-48 h were detected. TLR1,2,4,6,7,8,9,10 genes expression were detected in sheep alveolar macrophages except that of TLR3,5. α-tubulin and GAPDH have significant increase in LPS post stimulation and were not suitable as HKG. The specimen was normalized with cell number and total RNA. TLR2,4 mRNA expression rapidly increased post-stimulation and peaked at 20 min post-stimulation and this high level was maintained throughout the procedure. Sheep TLR2, 4 were sensitive in LPS-stimulated AM and participate in early stage of immune response.