The Correlation Of PI3K/Akt Cell Signaling Pathway On Progesterone Production Of Cultured Luteal Cells Of Goats

Immortalized in vitro proliferation and survival of luteal cells after adding a certain concentration of exogenous VEGF(Vascular Endothelial Growth Factor, VEGF) and VEGFR-2 receptor on the cell membrane after activation of PI3 K / Akt signaling pathway association between progesterone secretion and expose VEGF / VEGFR-2 / PI3 K / Akt signaling pathways in the luteal cell proliferation and survival in the role of progesterone secretion process undertaken.This test uses immortalized luteal cells and umbilical vein endothelial cells.Their quantity ratio is approximately 1: 1,and co-culture with MTT assay to test cell proliferation survival, and the use of cell samples ELISA kit double-antibody sandwich assay in pregnancy 3β-HSD, P450 scc and STAR1 mRNA expression change while the analysis of progesterone synthesis of three specific marker enzymes with real-time quantitative PCR(qRT-PCR).The obtained results were as follows:1.A certain concentration of exogenous VEGF(50ng / ml) on co-cultured cells have significant role in promoting the survival and proliferation.Co-cultured cell survival play a catalytic role in 24 h and 48 h two points in time, the performance of the difference was significant(P <0.01).2.With different concentrations of exogenous LY294002 and a certain concentration of exogenous VEGF combination group can significantly inhibit the proliferation of co-cultured cell survival, which in 24 h and 48 h time showed a significant difference(P <0.01), VEGF + LY294002(50μM) when administered in combination, can significantly inhibit the proliferation of co-cultured cell survival(P <0.001).3.A certain concentration of exogenous VEGF(50ng / ml) for individual cultured luteal cells proliferation and survival have no significant effect(P> 0.05); different concentrations of exogenous LY294002 inhibition of cultured alone luteal cells proliferation and survival after PI3 K activity had significant inhibitory effects(P <0.01).4.Different concentrations of exogenous LY294002 and a certain concentration of exogenous VEGF combination group proliferation of cultured alone luteal cell survival had no significant inhibition(P> 0.05).5.Exogenous inhibitor LY294002 group and LY294002 + VEGF added co-administered group in luteal cells cultured alone for the inhibitory effect of progesterone secretion produced significantly different exogenous inhibitor LY294002 addition group at 24 and 48 h when they cultured alone can luteal cells secrete progesterone produced significant inhibitory effect(P <0.001), but LY294002 + VEGF group of co-administered separately cultured luteal cells secrete progesterone had no significant inhibitory effect(P> 0.05).6.LY294002 + VEGF combination group produced significant inhibitory effect(P <0.001) on the co-cultured cells secrete progesterone.VEGF on luteal cells were co-cultured in a system of showing a promotion, but for luteal cells cultured alone had no significant promoting effect. PI3 K inhibitor LY294002 can effectively inhibit the secretion of progesterone, but in luteal cells cultured alone LY294002 + inhibition of VEGF in combination group on cell proliferation was not significant.