The Study Of Butaprost And Fluprostenol In Regulating Growth Factors Expression On Bovine Endometrial Epithelial Cells

Prostaglandins(PGs)performing a wide range of functions in mammals play important roles in secretion of a variety of reproductive related active substances.Prostaglandins E2(PGE2)and prostaglandins F2α(PGF2α)are abundant in the bovine endometrium.Recently,studies demonstrated PGE2 and PGF2α could promote human endometrial repair through regulating the expression of growth factors.Whether the PGs has the same effect on bovine endometrial repair has not yet been analyzed in detail.In this study,bovine endometrial epithelial cells have been used as research object.Discussed the cell signalig of PGE2 receptors agonists Butaprost and FP recptor agonist Fluprostenol in endometrium of bovine and the regulation function of growth factors related to tissue repair.Detailed research contents are as follows:(1)Culture bovine endomeitrial epithelial cells and analysis of PGs receptor.We cultured bovine endomeitrial epithelial cells combined enzyme digestion method and mechanical method and identified cells using immunofluorescence technique.Quantitative real-time PCR assays and Westernblot measured types of PGs receptor.The results show EP2 receptor and FP receptor exit in bovine endometrial epithelial cells and can be activation by it’s receptor agonist.(2)The effects of EP2 receptor agonists Butaprost on bovine endometrial epithelial cells.Quantitative real-time PCR assays measured the signal pathway of EP2 receptor agonists Butaprost on bovine endometrial epithelial cells.Quantitative real-time PCR assays and Westernblot measured the Butaprost on growth factors expression.We set different time points to study the regulation of Butaprost on VEGF,CTGF,TGF-β1 and IL-8 expression.Results showed Butaprost could activate PKA and ERK signaling pathway and promote COX-2,growth factors expression significantly.This enhancement disappered after join EP2 receptor antagonist AH6809.Butaprost could enhanced the expression of VEGF significantly and 8h most obviously.It also enhanced the expression of CTGF,24-48 h increased most significantly.Compared to m RNA expression,protein expression have certain hysteresis.Butaprost could promote TGF-β1 expression in 8-48 h and enhance the IL-8 expression from 2h to 16 h.(3)The effects of FP receptor agonists Fluprostenol on bovine endometrial epithelial cells.Quantitative real-time PCR assays measured the signal pathway of FP receptor agonists Fluprostenol on bovine endometrial epithelial cells.Quantitative real-time PCR assays and Western blot measured the Fluprostenol on growth factors expression.We set different time points to study the regulation of Fluprostenol on VEGF,CTGF,TGF-β1 and IL-8 expression.Results showed Fluprostenol could activate PKC and ERK signaling pathway and promote COX-2,growth factors expression significantly.This enhancement disappered after join FP receptor antagonist AL8810.Fluprostenol could enhanced the expression of VEGF significantly and 8h most obviously.After adding Fluprostenol stimulate the CTGF expression enhanced from 2h to 48 h.Fluprostenol could promoted TGF-β1 expression at 4-48 h and enhanced the IL-8 expression from 2h to 16 h.Conclusions: Successfully cultured bovine endometrial epithelial cells.Demonstrate EP2 receptor and FP receptor exit in bovine endometrial epithelial cells and can be activation by it’s receptor agonist.Clarify that EP2 receptor agonists Butaprost and FP receptor agonists Fluprostenol can promote the expression of EP2 receptor and FP receptor in bovine endometrial epithelial cells.EP2 receptor agonists Butaprost and FP receptor agonists Fluprostenol could regulate VEGF,CTGF,TGF-β1,IL-8 and COX-2 expression in bovine endometrial epithelial cells.EP2 receptor agonists Butaprost can activate the PKA and ERK pathway and FP receptor agonists Fluprostenol can activation of PKC and ERK pathway.