| Purpose:Prostaglandin is a kind of twenty-carbon Unsaturated fatty acids that metabolized arachidonic acid by cyclooxygenase and which is widely distributed in h uman and animal body. Prostanoids is a key regulatory factor in the process of animal re production, and it can affect many reproductive physiological processes. These experi ments were designed to explore the expression of Prostaglandin E2and F2α synthase PGES and PGFS mRNA in bovine oviductal epithelial cells by estrogen(E2), and ex plore the expression of cyclooxygenase1(COX-1), cyclooxygenase2(COX-2) as wel1as PGES and PGFS mRNA in bovine oviductal epithelial cells by the prostaglandi n EP receptor agonists butaprost and FP receptor agonists fluprostenol, as well as the e xpression of PGE2and PGF2a mRNA by the prostaglandin EP receptor agonists butapro st and FP receptor agonists fluprostenol.Method:1) Establishing bovine oviductal epithelial cells in vitro culturing syste m, and the cells were used for the experiment when they passaged to fourth-generat ion.2) Using RT-PCR determined and measured the expression of prostaglandin E2and F2a synthase PGES and PGFS mRNA level when different times (2h,4h,8h,16h,24h and48h) and different concentrations of estrogen (10-12-10-9mol/L) were added to the cells.3) Using RT-PCR determined and measured the expressio n of prostaglandin E2and F2a synthase PGES and PGFS mRNA level when differe nt concentrations of estrogen (10-12-10-9mol/L) were added to the bovine oviductal epithelial cells of the fourth generation respectively. The interference of endogenous p rostaglandinsin in epithelial cells was eliminated by adding indomethacin (10-5-10"6mol/L).4) Different concentrations of butaprost (10-9-10-5mol/L) and different cone entrations of fluprostenol (10-9-10-5mol/L) were added to the bovine oviductal epith elial cells of the fourth generation to explore the expression of cyclooxygenase1(C OX-1), cyclooxygenase2(COX-2) as well as PGES and PGFS mRNA level. The interfe rence of endogenous prostaglandinsin in epithelial cells was eliminated by adding indo methacin (10-5mol/L).5) Using ELISA determined the expression of PGE2、PGF2a level when estrogen (10-12mol/L) and different concentrations of butaprost and flupr ostenol were added to the bovine oviductal epithelial cells of the fourth generation24h respectively.Result:Frist, we successfully obtained cells for detecting prostaglandin E2and F2a synthase mRNA expression, and the fourth generations of cells were growed well. Se cond, compared with the control, the levels of PGES and PGFS were significantly i ncreased when adding E2(10-10mol/L) and E2(10-12mol/L) and reacting for4h a nd24h(P<0.01). Third, the levels of PGE and PGFS were significantly increase d from the control after eliminating the interference of endogenous prostaglandinsin in epithelial cells by indomethacin (10-5mol/L)(P＜0.01). Fourth, The expression of C OX-1was no significant difference from control, but the expression of COX-2was si gnificantly higher than the control, and the expression was positively correlated wit h concentration when butaprost (10-9-10-5mol/L) and fluprostenol (10-9-10-5mol/L) w ere added to the bovine oviductal epithelial cells and incubated for4h. The prosta glandin PGE2and PGF2α expression level was decreased from the control, and negati vely correlated with concentration. The expression of cyclooxygenase1(COX-1), cy clooxygenase2(COX-2) as well as PGES and PGFS levels were similar as only additi on of butaprost and fluprostenol after eliminating the interference of endogenous prost aglandins in epithelial cells by indomethacin. Fifth, The expression of PGE2and PGF2αwere positively correlated with the concentration when estrogen and different concentrations (10-9-10-5mol/L) of butaprost and fluprostenol were added and incuba ted for24h respectively.Conclusion:The experimental results showed thatestrogen can promote the expres sion of Prostaglandin mRNA in oviduct epithelial cells of bovine. PGE2and PGF2α has a positive feedback regulation to COX-2, but has a negative feedback regulation t o PGES and PGFS. In general, E2has a positive feedback regulation to the secretion of PGE2and PGF2α. |
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