Erickson G F, Ryan K J
Endocrinology. 1975 Jul;97(1):108-13. doi: 10.1210/endo-97-1-108.
The purpose of this study was to establish the effects of trophic hormones on the production of estrogens by rabbit granulosa cells. A pure population of these cells was isolated from preovulatory follicles (1-15 mm in diameter) of estrous rabbits, and cultured for 6 days with either one or a combination of the following hormones: androstenedione, Pergonal (LH/FSH), dibutyryl cyclic AMP (Bu2cAMP), prostaglandin F2alpha (PGF2alpha), or prostaglandin E2 (PGE2). The medium was collected every 2 days and progesterone (P), estrone (E1) and estradiol-17 beta (E2beta) were measured by radioimmunoassay. Granulosa cells cultured as controls (i.e., without exogenous trophic hormones) secreted P spontaneously and its secretion was stimulated 100 to 1,000 fold with LH/FSH and Bu2cAMP, but not with PGF2alpha or PGE2. Androstenedione, either alone or with trophic hormones had no apparent effect on the cytology of the granulosa cells or their ability to secrete P. In the absence of exogenous androstenedione, the cultures produced very small amounts of E1 or E2beta (smaller than 100 pg/ml), either spontaneously or in response to LH/FSH, B12cAMP, PGF2alpha, or PGE2. Incubating granulosa cells with exogenous androstenedione (1 mug/ml) resulted in a 30- to 150-fold increase in E2beta production, which was stimulated an additional 3- to 5-fold with LH/FSH and Bu2cAMP, but not with PGF2alpha or PGE2. In most cultures, E2beta production was restricted to the first 2 days in vitro. Bu2cAMP, however, maintained E2beta production at relatively high levels throughout the duration of the experiment, but there was a progressive decrease in its production. The production of E1 was only 5 percent of E2beta, but the pattern of secretion was similar for both estrogens. These results suggest that cyclic AMP could have a role in regulating the synthesis of estrogens by rabbit granulosa cells.
本研究的目的是确定营养激素对兔颗粒细胞雌激素产生的影响。从发情期兔子的排卵前卵泡（直径1 - 15毫米）中分离出纯的颗粒细胞群体，并与以下一种或几种激素组合培养6天：雄烯二酮、 Pergonal（促黄体生成素/促卵泡生成素）、二丁酰环磷腺苷（Bu2cAMP）、前列腺素F2α（PGF2α）或前列腺素E2（PGE2）。每2天收集培养基，通过放射免疫测定法测量孕酮（P）、雌酮（E1）和雌二醇-17β（E2β）。作为对照培养的颗粒细胞（即不添加外源性营养激素）自发分泌P，其分泌受到促黄体生成素/促卵泡生成素和Bu2cAMP刺激100至1000倍，但不受PGF2α或PGE2刺激。单独使用雄烯二酮或与营养激素一起使用，对颗粒细胞的细胞学或其分泌P的能力没有明显影响。在没有外源性雄烯二酮的情况下，培养物自发或对促黄体生成素/促卵泡生成素、B12cAMP、PGF2α或PGE2产生的E1或E2β量非常少（小于100 pg/ml）。用外源性雄烯二酮（1微克/毫升）孵育颗粒细胞导致E2β产生增加30至150倍，促黄体生成素/促卵泡生成素和Bu2cAMP使其额外增加3至5倍，但不受PGF2α或PGE2刺激。在大多数培养物中，E2β产生仅限于体外培养的前2天。然而，Bu2cAMP在整个实验过程中使E2β产生维持在相对较高水平，但产量逐渐下降。E1的产生仅为E2β的5%，但两种雌激素的分泌模式相似。这些结果表明环磷腺苷可能在调节兔颗粒细胞雌激素合成中起作用。
