The effect of LH/FSH, dibutyryl cyclic AMP, and prostaglandins on the production of estrogens by rabbit granulosa cells in vitro.
The effect of LH/FSH, dibutyryl cyclic AMP, and prostaglandins on the production of estrogens by rabbit granulosa cells in vitro.
复制标题
LH/FSH、二丁酰环腺苷酸和前列腺素对体外兔颗粒细胞产生雌激素的影响。
作者：
G. Erickson;K. Ryan
本研究的目的是建立营养激素对兔颗粒细胞产生雌激素的影响。从发情期家兔的排卵前卵泡（直径1-15 mm）中分离出这些细胞的纯群体，并与以下激素中的一种或组合培养6天：雄烯二酮、孕酮（LH/FSH）、二丁酰环AMP（Bu 2 cAMP）、前列腺素F2 α（PGF 2 α）或前列腺素E2（PGE 2）。每隔2天收集培养基，通过放射免疫法测定孕酮（P）、雌酮（E1）和雌二醇-17 β（E2 β）。作为对照培养的颗粒细胞（即，无外源性营养激素）自发分泌P，其分泌被LH/FSH和Bu 2 cAMP刺激100至1，000倍，但不被PGF 2 α或PGE 2刺激。雄烯二酮，无论是单独或与营养激素没有明显的影响颗粒细胞的细胞学或其分泌P的能力。在没有外源性雄烯二酮，培养物产生非常少量的E1或E2 β（小于100 pg/ml），无论是自发或响应LH/FSH，B12 cAMP，PGF 2 α，或PGE 2。将颗粒细胞与外源性雄烯二酮（1 μ g/ml）孵育，导致E2 β产生增加30- 150倍，LH/FSH和Bu 2cAMP可额外刺激E2 β产生3- 5倍，而PGF 2 α或PGE 2则不能。在大多数培养物中，E2 β的产生仅限于体外培养的前2天。然而，Bu 2cAMP在整个实验期间维持E2 β产生在相对高的水平，但其产生逐渐减少。E1的产量仅为E2 β的5%，但两种雌激素的分泌模式相似。这些结果表明，环磷酸腺苷可能有调节兔颗粒细胞的雌激素合成的作用。
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.