感染金黄色葡萄球菌小尾寒羊外周血白细胞和乳腺组织中TLRs基因的表达

Expression of the TLRs gene in peripheral white blood cell and mammary tissue of small-tail Han sheep infected with Staphylococcus aureus

【目的】Toll样受体是一种调节天然免疫反应的I型跨膜蛋白受体,研究拟通过检测TLRs在金黄色葡萄球菌引起的羊乳房炎发病过程中的表达情况,为进一步探索TLRs在绵羊乳房炎的作用机理奠定基础.【方法】以泌乳期的小尾寒羊为实验动物,运用实时荧光定量PCR方法检测TLR1、TLR2、TLR3、TLR4、TLR5基因在小尾寒羊外周血白细胞和乳腺组织中mRNA水平的相对表达量.【结果】TLR1、TLR2、TLR3、TLR4、TLR5在对照组和感染组的小尾寒羊外周血白细胞和乳腺组织中均可表达.TLR5在感染组的表达量低于对照组.在小尾寒羊外周血白细胞中,TLR2在12h的表达量较感染后的其他时间段高,并且12h的表达量较对照组显著升高;TLR1和TLR3的表达量与对照组相比显著降低;TLR4在感染后的表达量低于对照组.在小尾寒羊乳腺组织中,TLR2在感染36h的表达量较对照组显著增高,在72h降到最低,但其表达量仍高于对照组;TLR3在感染后36h和48h的表达量均低于对照组.TLR4除24h的表达量显著升高外,其余时间段均低于对照组.TLR1的表达量各时段无差异.【结论】TLRs参与了感染羊乳房炎早期的病理过程.

【Objective】Toll-like receptor is a type I transmembrane protein receptor that regulates the innate immune response.The study was conducted to explore the expression of TLRs in the pathogenesis of Staphylococcus aureus induced sheep mastitis,and to provide a foundation on the mechanism of action of TLRs in sheep mastitis for further study.【Method】Small-tail Han sheep in lactation period and infected with S.aureus was used as the experimental animal to detect the relative expression of TLR1,TLR2,TLR3,TLR4,TLR5 genes mRNA level in Small-tail Han sheep peripheral blood leukocyte and breast tissue by utilizing the method of real-time fluorescent quantitative PCR.【Result】The results showed that the TLR1,TLR2,TLR3,TLR4,TLR5 genes were expressed in peripheral blood leukocytes and breast tissue infected with S.aureus.The expression of TLR5 was lower than control.In peripheral blood leukocytes,the expression of TLR2 was higher than other treatment group after infected for 12 h.The expression of TLR2 gene was significantly higher in 12 h(P<0.05).The expression of TLR1 and TLR3 genes was significantly reduced(P<0.05)compared with control.The expression quantity of TLR4 gene was lower than control after infecting.In the breast tissue,the expression quantity of TLR2 gene in 36 h was increased significantly(P<0.05),and reached the lowest in 72 h,but the expression quantity was still higher than control.The expression quantity in infected 36 h and 48 h was higher than control.The expression quantity of TLR4 gene was lower than control,except in 24 h.The expression of TLR1 gene was not distinctly different from that of control.【Conclusion】TLRs involved in the early infection stage of sheep mastitis.