摘要
本文报道以氚标记Ｒ１８８１为特异性配基，直接测定男性包皮组织胞浆和核内雄激素受体（ＡＲ）的方法和结果。成年男性包皮组织直接制备胞浆；粗核沉淀经洗脱后用０．６ｍｏｌ／ＬＫＣｌ溶液抽提核浆。按Ｓｃａｔｃｈａｒｄ作图法分析了１１例样品及６份作为阳性对照的小鼠肝脏组织胞浆ＡＲ和核内ＡＲ。包皮组织胞浆ＡＲ最大受体结合容量（Ｂｍ）为２４．０３±７．１９ｆｍｏｌ／ｍｇ蛋白，解离常数（Ｋｄ）为１５．１３×１０－１０ｍｏｌ／ｌ；核内ＡＲ的Ｂｍ为２０４．３７±１１９．６２ｆｍｏｌ／ｍｇ蛋白，Ｋｄ为６．３８×１０－１０ｍｏｌ／Ｌ。３例完全型对雄激素不敏感综合征患者会阴皮肤活检组织的胞浆ＡＲ和核内ＡＲ均明显低于正常，核内ＡＲ的受体容量仅为６．１１～１１．２２ｆｍｏｌ／ｍｇ蛋白。由于不需经过组织和细胞培养，本方法更简便易行。
bjectives:To develop a method for measuring directly the cytosol and nuclear androgen receptor(AR)without tissue and cell culture.Design:A laboratory methodology and its clinical application. Setting:The Reproductive Endoerinology Laboratory of National Research Institute for Family Plan-ning. Suhjects:11 Chinese cdult men,3 patients with complete androgen insensitive Syndrome(CAIS)and 6 Intervention:Foreskin specimen were taken from the volunteer at the time of circumcision, the genital skin of patient were biopsied with his consent.Tissues were quickly frozen in liquid nitrogen and stored at -70℃. The fresh rat liver tissue’s were prepared at the day of receptor assay. Methods:About 0.5～1.0g tissues were homogenized directly and centrifuged at 12000 g to get cy-tosol.The nucleochylema were extracted with 0.6 mol/L KCl buffer from the nuclear pellets.Total protein was measured by Bradford’s method.Triamcinolone acetonide was used to reduce non-specific binding. The receptor radioassay(RRA ) was in the principle of‘Scatchard Plots’and calculated with the computer pro-gramme‘Symphony Release 2.0’.Results:The cytosol AR was chracterized by Bm 24.03± 7.19(x±s) fmol/mg protein and Kd 15.13×1O-10mol/L,and nuclear AR,Bm204.37±119. 62 mol/mg protein,Kd 6.38× 1O-10mol/L The capaci-ties of cytosol AR of 3 patients with CAIS were 2.29,O. 60 and 1. 35 fmol/mg protein;nuclear AR,13.22,7. 28 and 6.11 fmol/mg protein. The cytosol of rat livers, Bm 4. 43～5. 25 fmol/mg protein and Kd 7. 29～13. 80× 10-10mol/L,were good quality control. Conclusion:The method,without tissue and fibroblast culture, was more practical and easy to per-fom. The nuclear AR was of much higher level and higher affinity to ligand than the eytosol AR.The vol-umes of AR(espicially nuclear AR)of 3 patients with CAIS were much lower than those of normal male adults. It shown that these androgen resistances were caused by AR defect.
出处
《生殖医学杂志》
CAS
1995年第4期201-205,共5页
Journal of Reproductive Medicine
