| Bovine mastitis is a common disease in the dairy industry,causing enormous economic losses.As the most important pathogen of infectious mastitis,Staphylococcus aureus can invade bovine mammary epithelial cells,thereby evading immune defense and causing persistent infection.Recently,autophagy is considered to be an important mechanism for host cells to clear intracellular pathogens.Therefore,in this study,a model of Staphylococcus aureus infection on dairy mammary epithelial cells was established.From the perspective of intracellular pathogen survival and cell autophagy,the key protein p38?was selected.The activation level of P38?was detected by Western Blot to verify that The P38MAPK signaling pathway may be activated during the induced MAC-T cell autophagy of Staphylococcus aureus.Then,in order to further verify whether P38MAPK plays an important role in Staphylococcus aureus induced MAC-T cell autophagy,P38?inhibitor SB203580 and activator anisomycin were used to detect LC3 immunofluorescent positive aggregation points,while using Western Blot detects P62 and LC3 protein levels.The Western Blot detection of ULK1ser757er757 phosphorylation was then verified that p38?inhibits autophagy by phosphorylating the ULK1ser757 site,which affects the survival of S.aureus.The results showed that:After S.aureus infected MAC-T cells,the p38MAPK pathway was activated in a time-dependent manner.The addition of inhibitor SB203580 promoted autophagy flow caused by S.aureus infection with MAC-T,up-regulated the ratio of LC3 II/LC3 I and reduced the expression of autophagy-related protein P62 and the phosphorylation of ULK1ser757,After using anisomycin,it inhibited the autophagy flow caused by S.aureus infection with MAC-T,reduced the ratio of LC3 II/LC3 I,inhibited the degradation of autophagy-related protein P62,up-regulated the phosphorylation of ULK1ser757,and caused intracellular infection Staphylococcus aureus decreased... |
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