Provencher P, Lorrain A, Bélanger A, Fiet J
Laval University Medical Centre, MRC Group in Molecular Endocrinology, Sainte-Foy, Quebec, Canada.
J Steroid Biochem. 1990 Aug 28;36(6):589-96. doi: 10.1016/0022-4731(90)90177-t.
Steroidogenesis was studied in guinea-pig glomerulosa-fasciculata cells maintained in primary culture for up to 7 days. The basal secretion which remained stable for the first 2 days in culture rapidly rose to reach a plateau on day 4 at levels 6-7-fold higher than those observed during the first 2 days of culture while the maximal response to ACTH in terms of cortisol and androstenedione secretion was fairly stable throughout the 7-day period. Exposure of glomerulosa-fasciculata cells to ACTH caused a stimulation of pregnenolone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone, corticosterone, 11-deoxy-corticosterone, 11-deoxycortisol, cortisol, dehydroepiandrosterone, androstenedione, 11 beta-hydroxyandrostenedione and aldosterone while, after 48 h of incubation, a marked accumulation of end-products, namely cortisol and 11 beta-hydroxyandrostenedione, was observed. The half-maximal steroidogenic response to ACTH occurred at concentrations varying between 1.7 x 10(-11) and 1.1 x 10(-10) mol/l for the 12 steroids examined. Addition of 8-bromoadenosine 3', 5'-cyclic monophosphate stimulated steroid secretion in a dose-dependent manner. Maximal response to 8-bromoadenosine 3', 5'-cyclic monophosphate was obtained at 1 mmol/l, and no further rise of steroid secretion was observed after addition of ACTH. Incubation of glomerulosa-fasciculata cells with labeled corticosterone, cortisol and androstenedione indicates that only androstenedione can be converted into 11 beta-hydroxyandrostenedione, thus suggesting that this end-product is a good parameter of the C-19 steroid production by guinea-pig glomerulosa-fasciculata cells in primary culture. The present data confirm that guinea-pig glomerulosa-fasciculata cells in primary culture provide an interesting model for the study of the regulation of C-19 steroid formation by the adrenals.
在原代培养长达7天的豚鼠肾小球-束状带细胞中研究了类固醇生成。基础分泌在培养的前两天保持稳定，在第4天迅速上升至平台期，其水平比培养的前两天观察到的水平高6 - 7倍，而在整个7天期间，促肾上腺皮质激素（ACTH）对皮质醇和雄烯二酮分泌的最大反应相当稳定。将肾小球-束状带细胞暴露于ACTH会刺激孕烯醇酮、17-羟孕烯醇酮、孕酮、17-羟孕酮、皮质酮、11-脱氧皮质酮、11-脱氧皮质醇、皮质醇、脱氢表雄酮、雄烯二酮、11β-羟雄烯二酮和醛固酮的分泌，而在孵育48小时后，观察到终产物即皮质醇和11β-羟雄烯二酮的明显积累。在所检测的12种类固醇中，对ACTH的半数最大类固醇生成反应发生在浓度介于1.7×10⁻¹¹和1.1×10⁻¹⁰mol/L之间。添加8-溴腺苷3',5'-环磷酸以剂量依赖方式刺激类固醇分泌。在1 mmol/L时获得对8-溴腺苷3',5'-环磷酸的最大反应，添加ACTH后未观察到类固醇分泌的进一步增加。用标记的皮质酮、皮质醇和雄烯二酮孵育肾小球-束状带细胞表明，只有雄烯二酮可以转化为11β-羟雄烯二酮，因此表明该终产物是原代培养的豚鼠肾小球-束状带细胞产生C-19类固醇的良好参数。目前的数据证实，原代培养的豚鼠肾小球-束状带细胞为研究肾上腺对C-19类固醇形成的调节提供了一个有趣的模型。
