Detection and Regulation of the Messenger for a Putative Bovine Endometrial 9-Keto-Prostaglandin E2 Reductase: Effect of Oxytocin and Interferon-Tau1
Detection and Regulation of the Messenger for a Putative Bovine Endometrial 9-Keto-Prostaglandin E2 Reductase: Effect of Oxytocin and Interferon-Tau1
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假定牛子宫内膜 9-酮-前列腺素 E2 还原酶信使的检测和调节：催产素和干扰素-Tau1 的作用
作者：
E. Asselin;M. Fortier
在生殖过程中，前列腺素（PG） E2 （PGE2）和PGF2α发挥着重要的作用，但它们往往发挥相反的作用。在体内识别妊娠时，PGF2α在反刍动物和包括人类在内的大多数物种中被认为是黄体溶解因子，而PGE2可能发挥黄体保护作用。我们之前已经证明，重组干扰素-tau (rIFN-τ)，负责识别反刍动物怀孕的胚胎信号，在体外刺激PGE2和前列腺素内过氧化物合成酶2 （Ptgs2，也称为环氧化酶2）基因在上皮和内膜基质细胞中的表达。由于PGE2是基质细胞产生的主要前列腺素，对Ptgs2的影响可以解释PGE2输出的增加。然而，在高浓度下，重组羊（ro） IFN-τ通过将原代PG从PGF2α转变为PGE2而作用于上皮细胞。产生的初级PG的这种变化可以解释为PGF合成酶（PGFS）活性的降低或PGE合成酶活性的增加，或者通过将PGE2转化为PGF2α的PGE2 - 9-酮还原酶的调节。因此，我们研究了PGFS和pge2 -9-酮还原酶（9K-PGR） mrna的调控，这两种酶导致PGF2α的产生。其他研究描述了9K-PGR在不同物种的子宫、卵巢、肾脏和肝脏中的活性，并证实该酶可能同时具有9K-PGR和20α-羟基类固醇脱氢酶（20α-HSD）活性。有人认为9K-PGR和20α-HSD是相同的酶。从已发表的兔20α-HSD/9K-PGR和大鼠20α-HSD cdna同源核苷酸序列中选择引物序列，通过逆转录-聚合酶链反应（RT-PCR）扩增出317碱基对（bp）片段，并进行克隆和测序。与家兔和大鼠20α-HSD的同源性分别为83%和78%。采用RT-PCR半定量研究20α-HSD/9K-PGR和前列腺素F合成酶（PGFS） mRNA在体外培养上皮细胞中的表达。用roIFN-t刺激细胞导致双相反应，低剂量（1 ng/ml）抑制PGF2α的产生，高剂量（10 μg/ml）刺激PGE2。PGE2的升高伴随着9K-PGR和PGFS mRNA基因表达的降低。我们还研究了催产素（OT）的作用，OT的存在对9K-PGR或PGFS基因表达均无影响。20α-HSD/9K-PGR转录本在牛其他组织（肝脏>肾脏>睾丸>卵巢）中也有不同强度的表达。我们认为9K-PGR和PGFS可能是围着床期子宫内膜特异性PGs调控的关键酶。
Abstract During reproductive processes, prostaglandin (PG) E2 (PGE2) and PGF2α play important roles in which they often exert opposite effects. At the time of recognition of pregnancy in vivo, PGF2α is recognized as the luteolytic factor in ruminants and in most species including human, whereas PGE2 may exert a luteoprotective action. We have previously demonstrated that recombinant interferon-tau (rIFN-τ), the embryonic signal responsible for recognition of pregnancy in ruminants, stimulated in vitro the production of PGE2 and prostaglandin-endoperoxide synthase 2 (Ptgs2; also called cyclooxygenase-2) gene expression in both epithelial and stromal endometrial cells. Since PGE2 is the major prostaglandin produced by stromal cells, the effect on Ptgs2 could explain the increase in PGE2 output. At high concentrations, however, recombinant ovine (ro) IFN-τ acts on epithelial cells by changing the primary PG produced from PGF2α to PGE2. This change in the primary PG produced could be explained by a decrease in PGF synthase (PGFS) activity or an increase in PGE synthase activity, or by modulation of a putative PGE2–9-ketoreductase, which converts PGE2 into PGF2α. Therefore, we have investigated the regulation of the mRNAs for PGFS and PGE2-9-ketoreductase (9K-PGR), two enzymes that lead to the production of PGF2α. Others have described 9K-PGR activity in uterus, ovaries, kidney, and liver of different species and have established that this enzyme could possess both 9K-PGR and 20α-hydroxysteroid dehydrogenase (20α-HSD) activity. Some have concluded that 9K-PGR and 20α-HSD are identical enzymes. Using primers sequences chosen from homologous nucleotide sequences of published rabbit 20α-HSD/9K-PGR and rat 20α-HSD cDNAs, a 317-base pair (bp) fragment was amplified by reverse transcription-polymerase chain reaction (RT-PCR), cloned, and sequenced. Homologies of 83% and 78% were found with rabbit and rat 20α-HSD, respectively. The presence of 20α-HSD/9K-PGR and prostaglandin F synthase (PGFS) mRNA expression was studied semiquantitatively in cultured epithelial cells using RT-PCR. Stimulation of cells with roIFN-t resulted in a biphasic response, an inhibition of PGF2α production at low dose (1 ng/ml) and a stimulation of PGE2 at high dose (10 μg/ml). The increase of PGE2 was accompanied by reduced 9K-PGR and PGFS mRNA gene expression. The effect of oxytocin (OT) was also studied, and the presence of OT had no effect on either 9K-PGR or PGFS gene expression. The 20α-HSD/9K-PGR transcript was also detected in other bovine tissues at different intensity (liver > kidney > testis > ovaries). We believe that the 9K-PGR and PGFS can be key enzymes in the regulation of specific PGs in the endometrium during the periimplantation period.
影响因子：
11.2
作者：
C. Pontzer;F. Bazer;Howard M. Johnson
通讯作者：
C. Pontzer;F. Bazer;Howard M. Johnson
鉴定主要催乳素调节蛋白 20 α-羟基类固醇脱氢酶：协调调节其活性、蛋白含量和信使核糖核酸表达。
影响因子：
4.8
作者：
Albarracin,CT;Parmer,TG;Duan,WR;Nelson,SE;Gibori,G
通讯作者：
Gibori,G
DOI：
10.1016/s0006-291x(88)80109-8
发表时间：
1988-04-29
影响因子：
3.1
作者：
PONTZER, CH;TORRES, BA;JOHNSON, HM
通讯作者：
JOHNSON, HM