Study On The Expression Of SBD-1 In Ovine Ruminal Epithelial Cells Induced By Crude Extract Of Saccharomyces Cerevisiae Cell Wall Protein

The current damage caused by antibiotic abuse to human and animal health and the damage to the environment is increasingly prominent,forcing researchers to constantly look for substances that can replace antibiotics.Defensins,which have unlimited potential as alternative antibiotics,are a class of small-molecule antimicrobial peptides(AMPs)with broad-spectrum antimicrobial activity.Beta-defensins are the most widely distributed class of defensins in the family of defensins,widely distributed in mammals,birds and fish,while sheep beta-defensin-1(SBD-1)is the most widely distributed defensin in the gastrointestinal tract of sheep,especially in the rumen of sheep.This study investigated the effects of crude extracts of Saccharomyces cerevisiae cell wall proteins on the expression of SBD-1 in ovine ruminal epithelia cells(ORECs)at mRNA and protein levels.The second step is to disrupt the cultured Saccharomyces cerevisiae by ultrasonication to extract the cell wall,and then extract the crude extracts of Saccharomyces cerevisiae cell wall proteins by complex enzymatic method.The third step stimulated ORECs at 0,12.5,25,50 and 100 μg/mL with crude extracts of Saccharomyces cerevisiae cell wall proteins concentration for 2,4,8 and 12 h.Finally,quantitative real-time PCR(qPCR)and enzyme-linked immunosorbent assay(ELISA)were used to detect different concentrations with crude extracts of Saccharomyces cerevisiae cell wall proteins to induce SBD-1 mRNA at different time points in ORECs.Results:In this experiment,the ORECs were successfully cultured in vitro,and when the primary ORECs grow to the twelfth day,the cells basically fill the bottom of the cell culture flask.The crude extracts of Saccharomyces cerevisiae cell wall proteins extracted by the composite enzyme method was detected by SDS-PAGE,and the results satisfied the color development range.When crude extracts of Saccharomyces cerevisiae cell wall proteins concentration was 100 μg/mL and induced ORECs for 12 h,the expression level of SBD-1 was maximized.