Distribution of Toll-like Receptors(TLRs) and TLR2,TLR4 Expression During LPS Stimulus Sheep Alveolar Macrophage

doi:10.7668/hbnxb.2010.01.006

Abstract

Abstract: 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-PCR) was used to detect the distribution of sheep TLR1-TLR10 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.

Key words: Sheep, Alveolar macrophage, LPS, Toll like receptor, Real-time PCR

CLC Number:

S852.4+2

SHENG Jin-liang1,CHEN Chuang-fu1,YANG Xia1,WANG Yuan-zhi2,ZHANG Hui1. Distribution of Toll-like Receptors(TLRs) and TLR2,TLR4 Expression During LPS Stimulus Sheep Alveolar Macrophage[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(1): 30-35. doi: 10.7668/hbnxb.2010.01.006.

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