Effects Of Matrine On The Adhesion Of Staphylococcus Aureus To Bovine Mammary Epithelial Cells

Bovine mammary epithelial cells(BMECs)are exposed to pathogens,and their interaction with pathogenic microorganisms has a very important role for the development of the disease process.Adhesion is one of the prerequisites for pathogens to invade the host cells,and the pathogen first adheres to the host cells and then colonizes the host cells to play a pathogenic role.In the process of bovine mastitis,adhesion is the key step for pathogens infected breasts.This study is to investigate the effect of matrine on the adhesion of Staphylococcus aureus to BMECs in order to provide a theoretical basis for explaining the characteristics of pathogens adhesion to BMECs and the mechanism of pathogen invasion of BMECs.First,Tissue mass cell culture method and milk separation method were used to isolate and culture cells,then BMECs were purified and identified.The effects of different time,pH and temperature on the adhesion of BMECs to Staphylococcus aureus were studied.Different concentrations of matrine were added into the cell culture medium to culture BMECs.The effects of matrine on the proliferation and apoptosis of BMECs were detected.The effects of matrine on the proliferation,apoptosis and antioxidant capacity of BMECs were evaluated comprehensively to select matrine levels that was able to promote the proliferation of BMECs and enhance the antioxidant capacity of BMECs.The effects of matrine on the adhesion were studied by using resveratrol,dexamethasone and alfalfa flavonoids as control groups.The results showed that tissue mass cell culture method and milk separation method were able to culture BMECs.The karyotype was normal(2n = 60,XX)and keratin was positive,which indicated that BMECs were successfully established in vitro.Under a microscope,P4?P10 after purification were polygonal or droplet type,the morphology of P10-P14 changed to form a clear "mesh"structure.Their growth curves of P4-P14 were "S" type,which met the general laws of biological growth;but P12 and P14 growed relatively slow.After staining with Wright-Giemsa,it was found that vacuolar structure was appeared between P12 and P14.The optimum time for adhesion was 2.5 h,the optimum pH was between 7.0 and 7.5,and the optimum temperature was 37?.By detecting the effect of matrine on the proliferation and antioxidant capacity of BMECs,it was found that matrine(50 ?g/mL)was able to promote BMECs proliferation and increase the antioxidant capacity.Matrine(50 ?g/mL)was used to adhesion experiment,and it was found that compared with resveratrol,dexamethasone and alfalfa flavonoids,the effect of matrine on the adhesion had a significant inhibitory.The results of the above studies revealed that the purified cells were BMECs,and the traits of P4?P10 were more stable and used for subsequent experiments.The ptimum time for Staphylococcus aureus to adhere to BMECs was 2.5 h and the optimum pH was 7.0 to 7.5,The optimum temperature was 37 ?,and matrine(50 ?g/mL)had a significant inhibitory effect on adhesion.