Sterling K, Tsuboyama G, Brenner M A
Department of Medicine, Columbia University College of Physicians and Surgeons, New York, NY.
Endocr Res. 1988;14(1):109-16. doi: 10.1080/07435808809036343.
Isolated dispersed rat liver cells were prepared by hypothyroid Sprague-Dawley rats. The cells were incubated under 95% O2/5% CO2 in Krebs-Ringer-bicarbonate buffer at pH 7.3-7.4 at 37 degrees C. The medium had been enriched with 2% bovine serum albumin (previously stripped of thyroid hormone) and 5-10 mM alanine as substrate. Two hour incubations were carried out with or without added triiodothyronine (T3) at 3 nM or 300-1,000 nM concentrations. Oxygen consumption determined at the end of the period of incubation with the Clark oxygen electrode showed stimulation above control values in the hormone treated flasks; parallel studies in which cycloheximide (100 microM) had been added to cells to block protein synthesis also showed enhanced oxygen consumption in response to T3. The results indicated a response to the hormone not dependent on new protein formation.
通过甲状腺功能减退的斯普拉格-道利大鼠制备分离的分散大鼠肝细胞。细胞在37℃下于pH 7.3 - 7.4的 Krebs-林格碳酸氢盐缓冲液中，在95% O₂/5% CO₂条件下孵育。培养基中添加了2%牛血清白蛋白（预先去除甲状腺激素）和5 - 10 mM丙氨酸作为底物。在添加或不添加浓度为3 nM或300 - 1000 nM的三碘甲状腺原氨酸（T3）的情况下进行两小时孵育。在孵育期结束时用克拉克氧电极测定的耗氧量显示，激素处理的培养瓶中的耗氧量高于对照值；在细胞中添加环己酰亚胺（100 μM）以阻断蛋白质合成的平行研究也显示，对T3的反应耗氧量增加。结果表明对该激素的反应不依赖于新蛋白质的形成。
