17beta-雌二醇通过beta受体和cAMP-ERK1/2级联调节仔猪睾丸支持细胞cyclinA2 mRNA的表达

17beta-Estradiol Regulates the Expression of CyclinA2 mRNA of Cultured Immature Boar Sertoli Cells via Estrogen Receptor beta,cAMP-PKA and ERK1/2

【目的】确定雌激素是否通过雌激素受体以及在cAMP-细胞外调节的蛋白激酶(ERK1/2)调节培养条件下,未成熟仔猪睾丸支持细胞中cyclinA2 mRNA的表达。【方法】以培养的仔猪睾丸支持细胞为试验材料,通过添加雌激素受体抑制剂以及各种信号通路的抑制剂,应用实时荧光定量PCR检测cyclinA2 mRNA的相对表达量。【结果】17beta-雌二醇(10-9 mol.L-1)以时间依赖的方式促进了cyclinA2 mRNA的表达(P<0.05),这一作用在30 min时到达到高峰。雌激素非特异性受体抑制剂ICI182780、雌激素受体beta抑制剂(ERbetaAnt)和雌激素受体alpha抑制剂(ERalphaAnt)单独作用对cyclinA2 mRNA的表达与空白对照相比没有显著影响(P>0.05),但ICI 182780与ERbetaAnt,而不是ERalphaAnt抑制了17beta-雌二醇诱导的cyclinA2 mRNA的表达(P<0.05)。17beta-雌二醇(10-9mol.L-1)和forskolin均促进了cyclinA2 mRNA的表达(P<0.05),而Rp-cAMP、H-89和U0126都抑制了17beta-雌二醇(10-9mol.L-1)的活性(P<0.05),但3种抑制剂单独作用时对cyclinA2 mRNA的表达与空白相比没有显著影响(P>0.05)。【结论】17beta-雌二醇主要通过ERbeta受体、影响cAMP的产生和ERK1/2激活,进而调节cyclinA2 mRNA的表达。

【Objective】 The objective of this study was to identify whether 17beta-estradiol regulates the expression of cCyclinA2 mRNA via the estrogen receptor beta（ERbetβa） and the cAMP-PKA-extracellular signal-regulated kinase（ERK1/2） pathway.【Method】Cultured immature boar sertoli cells were treated with 10-9 mol？L-1 17beta-estradiol,and real-time PCR was used to detect the expression of cyclinA2 mRNA.【Result】Treatment with 17beta-estradiol increased the expression of cyclinA2 mRNA from 15 min to 90 min（P〈0.05）.The effects of 17beta-estradiol activity peaked at 30 min compared to the control cells（P〈0.05）.Combined treatment with ICI182780 and ERβ reduced the 17beta-estradiol-induced increase in the expression of cyclinA2 mRNA（P〈0.05）,but ERβ alone did not significantly affect these parameters（P〈0.05）.Both 17beta-estradiol and forskolin induced the abundance of cyclinA2 mRNA（P〈0.05 for both）.Combined treatment with Rp-cAMP,H-89 and U0126 reduced the 17beta-estradiol-induced expression of cyclinA2 mRNA（P〈0.05 for all）,but Rp-cAMP,H-89 and U0126 alone had no significant effect on the abundance of cyclinA2 mRNA,compared to the control group.【Conclusion】 This study showed that 17beta-estradiol regulates the expression of cyclinA2 mRNA via the activation of ERβ,cAMP-PKA and ERK1/2.