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

 【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。

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

【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.