Studies On Secretion Of PGE₂and PGF₂。Induced By LPS In Bovine Oviduct Epithelial Cell

Prostaglandin E2is an important mediators of inflammation, which is a kind of eicosenoic unsaturated fatty acid produced by arachidonicacid through catalyzing of cyclooxygenase. When the body was suffered from bacterialtoxin, inflammatory mediators including prostaglandins E2were generated in cells. In this study, bacterial lipopolysaccharide was used in stimulation of bovine oviductal epithelial cells to detected synthetase expression that related to PGE2and PGF2α, and prostaglandin E2and F2α secretion influence. To clarify the pathological mechanism of excessively secreting prostaglandin E2and F2α when G-pathogen infection in bovine oviduct, and to collect basic experimental data for the treatment illness of oviductal secretion disorder caused by inflammation.Methods:1)Establish the method of bovine oviduct epithelial cells culture in vitro.2)Use different concentrations of LPS (1μg/mL,2.5μg/mL,5μg/mL,7.5μg/mL,10μg/mL) to treat the bovine oviductal epithelial cells after24h, by detecting the expression of COX-1,COX-2mRNA expression by real time PCR methods, to determine the optimal concentration of LPS.3) Real-time PCR were used to detect COX-1, COX-2, mPGES-1, mPGES-2, cPGES and PGFS mRNA expression in bovine oviduct epithelial cells after LPS treatment at indicated time points (2h,4h,8h,16h,24h,48h,72h).4)Using COX inhibitors, NF-κB inhibitors and LPS treated the bovine oviduct epithelial cells, to detect prostaglandin synthase expression by real-time PCR.5) PGE2and PGF2a secretion in bovine oviduct epithelial cell were detected by ELISA after5μg/mL LPS treatment at different indicated time points(2h,4h,8h,16h,24h,48h,72h) and different inhibitorses; to analyze the effect of LPS on the ratio of production PGE2/PGF2α in bovine oviduct epithelial cell.Results:Firstly, succeeded in the culture of bovine oviductal primary and passage epithelial cells. Secondly, the expression of COX-1and COX-2were significantly increased in a concentration-dependent manner, after treatment with LPS (5μg/mL). Thirdly, after treatment with5μg/mL LPS, COX-1mRNA expression and COX-2mRNA expression reached maxlmum at16h and2h respectively; the levels of mPGES-1and mPGES-2mRNA expression were increased in16～72h; cPGES and PGFS mRNA expression significantly increased in4-72h and8-16h respectively. Fourthly, the ratio of production PGE2/PGF2α in bovine oviduct epithelial cell was changed after treatment with5μg/mL LPS. Finaly, expression of prostaglandin synthase,and secretion of PGE2and PGF2α were inhibited by Using COX inhibitors, NF-κB inhibitors and LPS treated the bovine oviduct epithelial cells.Conclusion:LPS regulate the mRNA expression of COX-1, COX-2, mPGES-1, mPGES-2, cPGES and PGFS, and secretion of PGE2and PGF2a though NF-κB path. Furthermore, endogenous prostaglandins play an important role in this regulation. LPS cause imbalance of PGE3/PGF2α,then affect normal oviduct microenvironment.