Chair of Molecular Cell Physiology and Endocrinology, Institute for Zoology, Technische Universität Dresden, Germany.
Mol Cell Endocrinol. 2010 Jan 15;314(1):41-52. doi: 10.1016/j.mce.2009.07.032. Epub 2009 Aug 15.
We have scrutinized the effects of the phytoestrogen genistein and three synthetic estrogen receptor agonists, 17 alpha-ethynylestradiol (EE), propylpyrazole-triol (PPT) and diarylpropionitrile (DPN) in the completely estrogen-free background of aromatase knockout (ArKO) mice by means of two routes of substance administration: oral via diet (per os; po) or subcutaneous injection (sc) with the intention to evaluate the ArKO mice as sensitive model organism for uterotrophic assays. Additionally, we were aiming to qualitatively analyze effects resulting from oral administration path, in particular for PPT and DPN. Therefore, we analyzed the resulting uterine wet weights (UWW) and epithelial heights as physiological endpoints of function as well as the gonadotropin levels. Moreover, the gene expression profiles of estrogen receptors as well as important uterine and ovarian estrogen-response genes were investigated by real-time PCR. The uterus of ArKO mice responded very sensitive upon the substitution with EE (sc 5 microg/kg BW; po 50 microg/kg BW) in a proliferative manner. This was evaluated inter alia by increased UWW and by up-regulation of the expression of proliferation-associated and estrogen-response genes. It is important to note, that ER alpha and ER beta-agonist, PPT and DPN respectively (po 5mg/kg BW and sc 0.5mg/kg BW), have only been used for sc applications so far. Here, effects resulting from oral application were qualitatively described and evaluated for their applicability. The UWW and expression of proliferation-associated genes were increased following both po and sc treatment with PPT. In contrast, DPN did not exert an increase of the UWW, but a significant decrease of proliferation-associated gene and protein expression. Additionally, a substantial hypoplasia was detectable in the uterine cross-sections of DPN-treated mice. On the other hand, the phytoestrogen genistein (sc 10mg/kg BW; po 70 mg/kg BW) did not cause detectable uterotrophic responses or large changes of uterine and ovarian gene expression profiles under the applied experimental conditions, but significantly reduced the elevated gonadotropin levels of ArKO mice. In summary, we showed the utility of ArKO mice to detect ER-specific effects, in particular those of PPT and DPN also when applied orally.
我们研究了植物雌激素金雀异黄素以及三种合成雌激素受体激动剂，17α-乙炔基雌二醇（EE）、丙基吡唑三醇（PPT）和二芳基丙腈（DPN）在芳香酶敲除（ArKO）小鼠完全缺乏雌激素的背景下通过两种途径给药的效果：经口途径（口服；po）或皮下注射（sc），目的是评估 ArKO 小鼠作为子宫增重分析的敏感模型。此外，我们旨在定性分析口服途径给药的效果，特别是针对 PPT 和 DPN。因此，我们分析了由此产生的子宫湿重（UWW）和上皮高度作为功能的生理终点，以及促性腺激素水平。此外，通过实时 PCR 研究了雌激素受体的基因表达谱以及重要的子宫和卵巢雌激素反应基因。ArKO 小鼠的子宫对 EE 的替代（sc 5μg/kg BW；po 50μg/kg BW）非常敏感，呈增殖性反应。这尤其可以通过增加 UWW 和上调增殖相关和雌激素反应基因的表达来评估。需要注意的是，ERα和 ERβ激动剂，分别为 PPT 和 DPN（po 5mg/kg BW 和 sc 0.5mg/kg BW），迄今为止仅用于 sc 应用。在这里，我们定性描述了口服应用的效果，并评估了其适用性。经口和 sc 应用 PPT 后，UWW 和增殖相关基因的表达均增加。相比之下，DPN 没有增加 UWW，但显著降低了增殖相关基因和蛋白的表达。此外，在 DPN 处理的小鼠的子宫横切面上可检测到明显的发育不良。另一方面，植物雌激素金雀异黄素（sc 10mg/kg BW；po 70mg/kg BW）在应用的实验条件下不会引起可检测的子宫增重反应或子宫和卵巢基因表达谱的大变化，但显著降低了 ArKO 小鼠升高的促性腺激素水平。总之，我们表明 ArKO 小鼠可用于检测 ER 特异性效应，特别是当以口服方式应用时 PPT 和 DPN 的效应。
