FRBI对CA125、EGF、E2和IP3分泌及ERβmRNA和蛋白表达的影响

Effect of FRBI on Secretion of CA125,EGF,E2 and IP3 and Expression of ERβ mRNA and Protein

为了探究促卵泡激素受体结合抑制剂(FRBI)是否影响卵母细胞中糖类抗原125(CA125)、表皮生长因子(EGF)、雌二醇(E2)、三磷酸肌醇(IP3)和cAMP分泌,雌激素受体β(ERβ)mRNA和蛋白表达水平,并阐明FRBI作用的信号通路。以绵羊卵丘-卵母细胞复合体(COCs)作为研究对象,在培养基中分别添加10,20,30,40μg/mL的FRBI,分别检测培养液中CA125、EGF、E2、IP3和cAMP的浓度,研究卵母细胞中ERβmRNA和蛋白表达水平。结果显示与对照组(CG)相比较,FRBI-3组和FRBI-4组CA125和IP3浓度降低。FRBI-1组CA125和FRBI-2组IP3浓度明显高于CG。E2浓度随FRBI添加量增加(从10μg/mL到30μg/mL)而升高,FRBI-3组E2浓度最高。FRBI-3组和FRBI-4组cAMP浓度明显低于CG和FSH组。FRBI组ERβmRNA和蛋白表达水平逐渐降低,FRBI-4组ERβ蛋白水平低于FSH(P<0.05)。FRBI浓度与EGF和E2呈正相关。结果表明,高剂量的FRBI能抑制绵羊卵母细胞中CA125、cAMP和IP3产生,增加E2浓度,降低卵母细胞ERβ基因和蛋白表达水平,FRBI可能抑制IP3和cAMP信号通路。

This study aimed at the investigation of the effects of FRBI on CA125,epidermal growth factor(EGF)Estradiol(E2),inositol triphosphate(IP3)and cAMP of secretion,expression of the ERβmRNA and protein in oocytes,also the elucidation of the signaling pathway of FRBI.Sheep cumulus-oocyte complexes(COCs)were taken as the experimental objects.PCR,western blot and ELISA were utilized to detect the expression levels of ERβmRNA and protein in COCs,and the concentrations of CA125 and EGF in the medium fluid,respectively.The concentrations of CA125 and IP3 in the RBI-3 and FRBI-4 groups reduced slightly in comparison with those in CG.E2 concentrations in the FRBI groups increased gradually as the FRBI doses rose from 10μg/mL to 30 mg/mL,with a maximum increment of the FRBI-3 group.The cAMP concentrations in the FRBI-3 and FRBI-4 groups decreased slightly compared to those CG and in the FSH group.The expression levels of ERβmRNA and protein decrease dgradually.The level of ERβprotein in the FRBI-4 group was less than that in the FSH group(P<0.05).FRBI had positive correlation with EGF and E2 concentrations.But,FRBI doses correlated negatively with the levels of cAMP and ERβmRNA and protein.These results revealed that higher doses of FRBI(30 or 40μg/mL)suppressed the production of CA125,cAMP and IP3,but promoted E2 production of sheep oocytes.FRBI treatment could decrease the expressions of ERβmRNA and protein in sheep oocytes.FRBI probably suppressed the signaling cascades of IP3 and cAMP.