Markoff E, Lee D W, Culler F L, Jones K L, Lewis U J
J Clin Endocrinol Metab. 1986 Apr;62(4):664-9. doi: 10.1210/jcem-62-4-664.
Using a combination of polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and immunoblotting with antihuman GH (anti-hGH) serum, we quantitatively and independently measured the major 22,000-dalton form of hGH (hGH22K) and the 20,000-dalton form (hGH20K). This technique was equally effective in assaying cell culture media or human sera. We studied isolated human pituitary cell cultures during a 16-day incubation period with and without stimulation by GH-releasing hormone (GHRH). Under basal conditions, the cells released 2.83 +/- 0.24 (+/- SEM) microgram hGH22K/ml . day and 0.67 +/- 0.17 microgram hGH20K/ml . day. GHRH (10(-8) M) treatment resulted in stimulation of both hGH22K and hGH20K by 24 h. We also measured both hGH22K and hGH20K in the sera of normal subjects before and after an iv bolus injection of 100 micrograms GHRH. hGH20K increased as did hGH22K. Peak concentrations of both variants occurred 45 min after GHRH administration. The results of this study indicate that the combination of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting is an accurate and effective means of separately assaying hGH22K and hGH20K. We also demonstrated that primary monolayer cultures of human pituitary cells are an excellent model system for study of the secretion of these two hGH variants.
我们运用十二烷基硫酸钠存在下的聚丙烯酰胺凝胶电泳与抗人生长激素（抗hGH）血清免疫印迹相结合的方法，对主要的22,000道尔顿形式的hGH（hGH22K）和20,000道尔顿形式（hGH20K）进行了定量和独立测量。该技术在检测细胞培养基或人血清方面同样有效。我们研究了分离的人垂体细胞培养物在16天培养期内有无生长激素释放激素（GHRH）刺激的情况。在基础条件下，细胞释放2.83±0.24（±SEM）微克hGH22K/毫升·天和0.67±0.17微克hGH20K/毫升·天。GHRH（10⁻⁸ M）处理在24小时时导致hGH22K和hGH20K均受到刺激。我们还测量了静脉推注100微克GHRH前后正常受试者血清中的hGH22K和hGH20K。hGH20K和hGH22K一样增加。两种变体的峰值浓度在给予GHRH后45分钟出现。本研究结果表明，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹相结合是分别检测hGH22K和hGH20K的准确有效方法。我们还证明，人垂体细胞的原代单层培养物是研究这两种hGH变体分泌的优良模型系统。
