Beyer H S, Carr F E, Mariash C N, Oppenheimer J H
Endocrinology. 1985 Jun;116(6):2669-76. doi: 10.1210/endo-116-6-2669.
We have previously established the value of 2-dimensional electrophoretic mRNA activity profiles for investigating the hepatic genomic response to several metabolic perturbations, such as thyroid hormone or GH treatment, diabetes, high carbohydrate diet, starvation, and uremia. We now report the effects of adrenalectomy and dexamethasone treatment, and compare these with alterations due to thyroidectomy and T3 treatment. Total rat hepatic RNA was isolated and translated in a reticulocyte lysate system. The [35S]methionine-labeled translated products were separated by 2-dimensional gel electrophoresis and quantified with computerized videodensitometry. Of 200 consistently quantifiable products, 14 (7%) were altered by adrenalectomy and dexamethasone, including 4 products (46, 47, 56, and 57) which have not been observed to change in previous studies from this laboratory. Adrenalectomy increased 5 and decreased 2 products, whereas dexamethasone increased 1 and decreased 8 products. Two products maintained the same directional shift in the transitions form adrenalectomy to control and from control to the dexamethasone-treated state. Thyroidectomy and T3 altered 13 products. Thyroidectomy increased 2 and decreased 7 products, whereas T3 treatment increased 6 and decreased 3 products. Four products maintained the same directional shift in the transitions from thyroidectomy to control and from control to the T3-treated state. In all of the manipulations performed (adrenalectomy, thyroidectomy, dexamethasone treatment, and T3 treatment), a total of 20 separate products changed. One third were affected by alterations of both the steroidal and thyroidal states. However, when adrenalectomy and thyroidectomy were compared, only 7% of the shifts were concordant, whereas 30% of the shifts were concordant when treatment with dexamethasone and T3 were compared. These results demonstrate that the mRNA activity response is highly specific for each hormonal manipulation. In addition, unanticipated interrelationships between steroidal and thyroidal states were observed. In some, the presence of T3 appears necessary for the suppressive effect of dexamethasone. Others show that T3 appears to inhibit a stimulatory effect of dexamethasone. Specificity of response to dexamethasone is emphasized by the lack of response to vitamin D, deoxycorticosterone, and dihydrotestosterone and by a different response to estradiol from dexamethasone.
我们之前已经确定了二维电泳mRNA活性图谱在研究肝脏基因组对多种代谢扰动的反应中的价值，这些扰动包括甲状腺激素或生长激素治疗、糖尿病、高碳水化合物饮食、饥饿和尿毒症。我们现在报告肾上腺切除术和地塞米松治疗的效果，并将其与甲状腺切除术和T3治疗所引起的变化进行比较。分离出大鼠肝脏总RNA，并在网织红细胞裂解物系统中进行翻译。用[35S]甲硫氨酸标记的翻译产物通过二维凝胶电泳进行分离，并用计算机视频密度计进行定量。在200种可 consistently 定量的产物中，有14种（7%）受到肾上腺切除术和地塞米松的影响，其中包括4种产物（46、47、56和57），在本实验室之前的研究中未观察到它们发生变化。肾上腺切除术使5种产物增加，2种产物减少，而地塞米松使1种产物增加，8种产物减少。有2种产物在从肾上腺切除术到对照以及从对照到地塞米松处理状态的转变中保持相同的方向变化。甲状腺切除术和T3改变了13种产物。甲状腺切除术使2种产物增加，7种产物减少，而T3治疗使6种产物增加，3种产物减少。有4种产物在从甲状腺切除术到对照以及从对照到T3处理状态的转变中保持相同的方向变化。在所有进行的操作（肾上腺切除术、甲状腺切除术、地塞米松治疗和T3治疗）中，共有20种不同的产物发生了变化。其中三分之一受到甾体和甲状腺状态改变的影响。然而，当比较肾上腺切除术和甲状腺切除术时，只有7%的变化是一致的，而当比较地塞米松和T3治疗时，30%的变化是一致的。这些结果表明，mRNA活性反应对每种激素操作具有高度特异性。此外，还观察到甾体和甲状腺状态之间存在意想不到的相互关系。在某些情况下，T3的存在似乎是地塞米松抑制作用所必需的。其他情况表明，T3似乎抑制了地塞米松的刺激作用。对维生素D、脱氧皮质酮和二氢睾酮无反应以及对雌二醇与地塞米松有不同反应强调了对地塞米松反应的特异性。
