Mechanism of action of 2-hydroxyestradiol on steroidogenesis in ovarian granulosa cells: interactions with catecholamines and gonadotropins involve cyclic adenosine monophosphate.
Mechanism of action of 2-hydroxyestradiol on steroidogenesis in ovarian granulosa cells: interactions with catecholamines and gonadotropins involve cyclic adenosine monophosphate.
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2-羟基雌二醇对卵巢颗粒细胞类固醇生成的作用机制：与儿茶酚胺和促性腺激素的相互作用涉及环磷酸腺苷。
DOI：
10.1095/biolreprod40.1.87
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发表时间：
1989
影响因子：
3.6
通讯作者：
Hammond,JM
中科院分区：
文献类型：
--
作者：
Spicer,LJ;Hammond,JM
以前，我们已经表明，2-羟基雌二醇（2-OH-E2）协同catecbolamines，以提高孕酮生产的猪颗粒细胞在体外。本研究旨在确定2-OH-E2和儿茶酚胺的协同作用是否1）由促性腺激素调节，2）由儿茶酚胺特有，3）由环磷酸腺苷（cAMP）介导。将1 - 3-mm猪卵泡的未分化颗粒细胞在无血清培养基中培养6 - 9天。在颗粒细胞培养的第7天，用促卵泡激素（FSH）预处理3天加共处理4天与共处理4天相比，没有显著改变饱和浓度肾上腺素（EPI; 2 μ g/ml）刺激的孕酮产生，但显著降低了4 μ g/ml 2 - 0H-E2的作用。在培养的第3 - 7天，FSH或LH共处理4天，显著增强了2-OH-E2和雌二醇（E2）刺激的孕酮产生，但在培养的第7天测量时，EPI没有。孕酮生成在LH存在的情况下，EPI、2-OH-E2或EPI +2-OH-E2处理4天刺激的细胞增殖率（表示为"对照的-倍"）分别为1.4 ± 0.2、8.2 ± 2.2和10.7 ± 1.0（n = 5次实验），在FSH存在下分别为1.9 ± 0.1、7.8 ± 1.4和10.6 ± 1.8（n = 3次实验）。与E2相似，2-OH-E2显著增强cAMP类似物8-溴-cAMP（0.5mM）对孕酮产生的刺激作用。饱和浓度的2-OH-E_2和EPI对孕酮分泌的协同作用在8-溴-cAMP存在下未观察到。然而，在磷酸二酯酶抑制剂3-异丁基-1-甲基黄嘌呤（IBMX，0.5 mM）的存在下，2-OH-E2和EPI之间的协同作用持续存在。此外，cAMP积累超过24小时（培养基加细胞提取物）刺激EPI或FSH显着升高，在2-OH-E2的存在下，但不是E2。这些结果表明：（1）2-OH-E_2能显著增强LH和FSH刺激的孕酮分泌，（2）2-OH-E_2与促性腺激素的协同作用至少与2-OH-E_2与儿茶酚胺的协同作用一样强; 3）与用大鼠颗粒细胞的研究相反，用促性腺激素和儿茶酚胺预处理或同时处理猪颗粒细胞，充其量，对儿茶酚胺能反应的累加效应;和4）由EPI或FSH刺激的2-OH-E2（而不是E2）-增强的孕酮产生的主要位点在cAMP产生的水平上发挥作用。
Previously we have shown that 2-bydroxyestradiol (2-OH-E2) synergizes with catecbolamines to enhance progesterone production by porcine granulosa cells in vitro. The present studies were undertaken to determine if the synergistic effects of 2-OH-E2and catecbolamines were 1) modulated by gonadotropins, 2) unique to catecholamines, and 3) mediated by cyclic adenosine monophosphate (cAMP). Undifferentiated granulosa cells from 1- to 3-mm porcine follicles were cultured in serum free medium for periods of 6–9 days. A 3-day pretreatment plus a 4-day cotreatment versus a 4-day cotreatment of granulosa cell cultures with follicle-stimulating hormone (FSH) did not significantly alter progesterone production stimulated by a saturating concentration of epinephrine (EPI; 2 μg/m1) but significantly reduced the effect of 4 μg/ml 2-0H-E2on Day 7 of culture. Four-day cotreatment of either FSH or luteinizing hormone (LH) from Day 3 to 7 of culture dramatically enhanced progesterone production stimulated by 2-OH-E2and estradiol (E2) but not by EPI when measured on Day 7 of culture. Progesterone production (expressed as “-fold of controls”) stimulated by 4-day treatment of EPI, 2-OH-E2, or EPI-plus-2-OH-E2was 1.4 ± 0.2, 8.2 ± 2.2, and 10.7 ± 1.0, respectively, in the presence of LH (n = 5 experiments), and 1.9 ± 0.1, 7.8 ± 1.4, and 10.6 ± 1.8, respectively, in the presence of FSH (n = 3 experiments). Similar to E2, 2-OH-E2significantly enhanced the stimulating effect of the cAMP analog 8-bromo-cAMP (0.5 mM) on progesterone production. The synergism between saturating concentrations of 2-OH-E2and EPI on progesterone production was not observed in the presence of 8-bromo-cAMP. However, the synergism between 2-OH-E2and EPI persisted in the presence of the phospbodiesterase inhibitor 3-isobuty1-1-methylxanthine (IBMX, 0.5 mM). In addition, cAMP accumulation over 24 h (medium plus cell extracts) stimulated by EPI or by FSH was significantly elevated in the presence of 2-OH-E2but not E2. Collectively, these results suggest that 1) 2-OH-E2can dramatically enhance LH- and FSH-stimulated progesterone production; 2) the synergism between 2-OH-E2and gonadotropins was at least as great as that of the synergism between 2-OH-E2and catecbolamines; 3) in contrast to studies with rat granulosa cells, pretreatment or concurrent treatment of porcine granulosa cells with gonadotropins and catecbolamines exerts, at best, an additive effect on the catecholaminergic response; and 4) a major locus of 2-OH-E2(but not E2)-potentiated progesterone production stimulated by EPI or FSH is exerted at the level of the generation of cAMP.