Expression Of TLRs And CK In Breast Tissue And Peripheral Blood Of Small Tail Han Sheep Infected With Staphylococcus Aureus

In this experiment,Small Tail Han Sheep were used as experimental animals to study mastitis caused by Staphylococcus aureus and to explore the expression of TLRs and CK and the relationship between mastitis.Real-time fluorescence quantitative PCR(FQ RT-PCR)was used to detect the relative expression levels of TLRs in breast tissue and peripheral blood leukocytes normal and infected Staphylococcus aureus at 12 h,24 h,36 h,48 h and 72 h.Western-blot method was used to detect the expression of TLRs protein in breast tissues of Tail Han sheep infected at 12,24,36,48 and 72 h.Detected the concentration of TNF-??IFN-??IL-1??IL-16 in serum by ELISA.The results showed that: TLRs are differentially expressed in infected breast tissue and peripheral blood leukocytes,TLR8 was not expressed,the expression of TLR3 were lower than the normal control group.In the infected peripheral blood leukocytes,the expression of TLR2 was significantly increased at 12 h;the expression of TLR6 and TLR7 was significantly increased at 48 h;the expression levels of TLR1 and TLR4 were all lower than that of the control group,There was no difference in the expression levels of TLR9 at 12 h and 24 h and TLR5,10 at 12 h,the expression levels in the rest of the time were lower than those in the control group.Other TLRs were expressed except TLR8 and TLR9 in the infected breast tissue,the expression level of TLR2 was higher in each period than that in the control group.The expression levels of TLR2 were significantly increased at12 h,24 h and 36 h.The expression levels of TLR7 and TLR10 were significantly increased at24 h and 36 h.TLR4 was significantly increased at 24 h.There was no difference in the expression of TLR1 and TLR6(P>0.05).The expression of TLR4 protein in breast tissue,TLR4 was expressed in six time periods,and the expression level was significantly increased at 24 h.The ELISA method was used to test the expression of cytokines in serum.The experimental results showed that the expression of IFN-? in serum was significantly higher than that of the control group at 24 h,36 h and 48 h.The expression level of IL-1? in serum reached the highest at 36 h,and then showed a gradually decreasing trend.The expression of IL-16 in serum was significantly increased at 24 h and 36 h,reached its peak at 24 h,and then gradually decreased.Compared with the control group,all other time periods were significantly increased except 72 h.The expression of TNF-? peaked at 36 h.The expression of IL-1? and IL-16 are relatively high in serum after infected with Staphylococcus aureus,which showed fluctuating changes.TLR2/10 is positively correlated with TNF-?,IFN-?,IL-1? and IL-16.There is also a significant positive correlation among TLR4/7 and IFN-??IL-16.The results provide basic data for further revealing the pathogenesis of Staphylococcus aureus mastitis in sheep.