FSH和17β-雌二醇联合作用对Cyclin D1 mRNA和Cyclin E1 mRNA表达的影响

摘要

The objective of the study was to identify whether the combination of FSH and 17-betaestradiol regulated the mRNA expression of Cyclin D1 and Cyclin E1 in the cultured immature boar Sertoli cells.Cultured immature boar Sertoli cells were treated with the combination of 17-beta-estradiol (10-9 mol · L-1) and FSH (50 ng · mL-1) and added a variety of different signaling pathway inhibitors.And Real-time PCR was applied to detect the mRNA expression of Cyclin D1 and Cyclin E1.Compared to FSH (50 ng · mL-1) or 17-beta-estradio1 (10-9 mol · L-1)alone,the combination of FSH and 17-beta-estradiol had no significant effect on the mRNA expression of Cyclin D1 mRNA and Cyclin E1 (P＞0.05).In addition,Rp-cAMP (cAMP inhibitor),Verapamil (L-type Ca2+ ionic channel inhibitor) or U0126 (ERK1/2 inhibitor) alone had no significant effect on the mRNA expression of Cyclin D1 and Cyclin E1 in comparison to the control group (no FSH or17-beta-estradiol) (P＞0.05).However,three inhibitors could reduce the mRNA expression of Cyclin D1 and Cyclin E1 in a dose-dependent way (P＜0.05,for all) when compared to the combined FSH (50 ng · mL-1) and 17-beta-estradiol (10-9 mol · L-1).cAMP,Ca2+ and ERK1/2 were involved in the effect of the combination of FSH and 17-beta-estradiol regulating the mRNA expression of Cyclin D1 and Cyclin E1.%为了确定FSH和雌激素联合作用对培养条件下未成熟仔猪睾丸支持细胞中Cyclin D1 mRNA和Cyclin E1 mRNA表达的影响.以培养的仔猪睾丸支持细胞为研究材料,通过添加各种信号通路的抑制剂,应用实时荧光定量PCR检测Cyclin D1 mRNA和Cyclin E1 mRNA的相对表达量.FSH(50ng·mL-1)和17β雌二醇(10-9mol·L-1)联合作用时对Cyclin D1 mRNA和Cyclin E1 mRNA表达的影响与FSH或17β-雌二醇(10-9 mol·L-1)单独作用相比无显著影响(P＞0.05)；环磷酸腺苷抑制剂(Rp-cAMP)、L-Ca2+离子通道抑制剂(Verapamil)和ERK1/2抑制剂(U0126)单独作用时对Cyclin D1 mRNA和Cyclin E1 mRNA表达与空白对照组相比无显著影响(P＞0.05),但以剂量依赖的方式抑制了FSH(50ng·mL-1)与17β-雌二醇(10-9 mol·L-1)联合作用对Cyclin D1 mRNA和Cyclin E1 mRNA的表达(P＜0.05).FSH和17β-雌二醇联合作用时通过cAMP、Ca2+内流和ERK1/2途径参与了FSH和17β-雌二醇对Cyclin D1 mRNA和Cyclin E1 mRNA在细胞中的表达.