Oikarinen A, Oikarinen H, Meeker C A, Tan E M, Uitto J
Department of Dermatology, University of Oulu, Finland.
Acta Derm Venereol. 1987;67(6):461-8.
In the present study, we have determined the specific glucocorticoid receptors in cultured human skin fibroblasts with [3H]dexamethasone as the ligand. The whole-cell assay was employed for determination of glucocorticoid receptor densities and binding affinities in fibroblast cultures established either from 16 healthy control subjects, from 4 patients with active progressive systemic sclerosis (PSS), from 3 patients with keloids and 3 patients with diabetes mellitus. The receptor densities in PSS, keloid, diabetes and control fibroblasts were in the same range, the values being 6.3 +/- 4.9, 7.7 +/- 3.6, 5.3 +/- 1.3 and 7.9 +/- 6.2 fmol/micrograms DNA (mean +/- SD), respectively. In further studies, the cells were incubated with 10(-7) M dexamethasone for 4 or 9 days before the receptors were assayed. The specific binding of [3H]dexamethasone in steroid treated cultures was 62 and 13% of that observed in controls, suggesting down-regulation. In contrast, incubation of fibroblasts with 10(-5) M all-trans-retinoic acid did not alter the binding of [3H]dexamethasone, suggesting lack of pharmacologic interference at the receptor level.
在本研究中,我们以[3H]地塞米松作为配体,测定了培养的人皮肤成纤维细胞中的特异性糖皮质激素受体。采用全细胞分析法测定了分别来自16名健康对照者、4名活动性进行性系统性硬化症(PSS)患者、3名瘢痕疙瘩患者和3名糖尿病患者的成纤维细胞培养物中的糖皮质激素受体密度和结合亲和力。PSS、瘢痕疙瘩、糖尿病和成纤维细胞对照中的受体密度处于同一范围,其值分别为6.3±4.9、7.7±3.6、5.3±1.3和7.9±6.2 fmol/μg DNA(平均值±标准差)。在进一步的研究中,在测定受体之前,将细胞与10^(-7) M地塞米松孵育4天或9天。在经类固醇处理的培养物中,[3H]地塞米松的特异性结合为对照中观察到的特异性结合的62%和13%,提示存在下调。相反,将成纤维细胞与10^(-5) M全反式维甲酸孵育不会改变[3H]地塞米松的结合,提示在受体水平缺乏药理干扰。