Kako T
Nihon Naibunpi Gakkai Zasshi. 1975 Jan 20;51(1):48-55. doi: 10.1507/endocrine1927.51.1_48.
Isolated adrenal cells were obtained surgically from patients with primary aldosteronism, breast cancer, or Cushing's syndrome. They were prepared by the modification of Sayers method, and incubated at 37 degrees C for 2 hours under 95% O-2-5% CO-2, in the medium of calcium-free Krebs-Ringer bicarbonate buffer containing 0.2% glucose and 0.5% bovine serum albumin, to which various doses of calcium, ACTH, dibutyryl cyclic AMP or cycloheximide were added. Steroid production was measured by the method of Silber et al. In isolated normal adrenocortical cells, 11-OHCS was produced by calcium alone in the absence of ACTH or dibutyryl cyclic AMP, while it was not produced by ACTH alone without calcium. 11-OHCS production by ACTH was decreased in the high concentration of calcium (10.16 mM, 12.70 mM). Cycloheximide partially blocked an increase in 11-OHCS synthesis induced by calcium. These data suggest that adenyl cyclase of human adrenocortical cells may be stimulated by calcium alone, supporting the notion that calcium is a second messenger. The ratio of 11-OHCS production by calcium alone to that by dibutyryl cyclic AMP was higher in adenoma cells than in normal cells. This may account for the character of autonomic steroid production in adrenocortical adenoma cells.
通过手术从原发性醛固酮增多症、乳腺癌或库欣综合征患者身上获取分离的肾上腺细胞。它们采用赛耶斯方法的改良版进行制备,并在含0.2%葡萄糖和0.5%牛血清白蛋白的无钙克雷布斯-林格碳酸氢盐缓冲液培养基中,于95% O₂ - 5% CO₂条件下37℃孵育2小时,向其中添加不同剂量的钙、促肾上腺皮质激素(ACTH)、二丁酰环磷腺苷或环己酰亚胺。类固醇生成采用西尔伯等人的方法进行测定。在分离的正常肾上腺皮质细胞中,在无ACTH或二丁酰环磷腺苷的情况下,仅钙就能产生11-羟皮质类固醇(11-OHCS),而仅ACTH在无钙时则不产生11-OHCS。在高浓度钙(10.16 mM,12.70 mM)时,ACTH诱导的11-OHCS生成减少。环己酰亚胺部分阻断了钙诱导的11-OHCS合成增加。这些数据表明,人肾上腺皮质细胞的腺苷酸环化酶可能仅受钙刺激,支持钙是第二信使这一观点。腺瘤细胞中仅钙产生的11-OHCS与二丁酰环磷腺苷产生的11-OHCS之比高于正常细胞。这可能解释了肾上腺皮质腺瘤细胞中自主性类固醇生成的特点。