一种非同位素5α-还原酶抑制剂体外筛选模型的建立和应用  

Establishment and application of a non-isotope model in vitro for screening 5α-reductase inhibitor

作　　者：况刚[1] 邹文俊[1] 高小平 

机构地区：[1]成都中医药大学,成都610075 [2]成都地奥制药集团药物研究所,成都610041

出　　处：《中药药理与临床》2005年第4期61-64,共4页Pharmacology and Clinics of Chinese Materia Medica

摘　　要：目的建立一种非同位素5α-还原酶抑制剂体外筛选模型。方法从大鼠前列腺组织中提取物5α-还原酶,与底物睾酮(T)以及供氢体NADPH共同组成了一种微量反应体系。反应产物二氢睾酮(DHT)用酶免技术(EIA)测定,以产物生成量确定酶活性;对反应底物、NADPH、酶浓度以及反应最佳温度和时间进行了优化。结果当底物、NADPH、酶提取物浓度分别为0.25μM、1mM和0.3mg,37℃反应60分钟可获最大酶活性。特异性5α-还原酶抑制剂非那雄胺显著降低5α-还原酶催化产物DHT的生成。进一步对24种中药进行筛选,姜黄、夏枯草等中药提取物具有5α-还原酶抑制剂作用。结论本文建立的非同位素方法有效、可行,可作为一种体外模型筛选和研究5α-还原酶抑制剂。For establishing a non-isotope model in vitro for screening 5alpha-reductase inhibitor , the 5alpha-reductase abstracted from the prostates of male rats and testosterone as a substrate together with nieotinamide adenine dinueleotide phosphare （NADPH） as a hydrogen supplier, were composed of a minim reactive system. The production dihydrotestosterone （DHT） was measured by the enzyme immunoassay（EIA）. The activity of enzyme was determined by the quantity of production. We optimized the concentrations of substrate, NADPH enzyme, temperature as well as time. When the concentrations of substrate, NADPH, enzyme were 0.25μM, 1mM and 0.3mg respec tively, the maximum activity of enzyme was achieved at 37℃ after 60min reaction. We established a positive control of specific 5alpha-reductase inhibitor by finasteride. The result showed it obviously reduced the production of DHT, which indicated that the activity of 5alpha- reductase was inhibited. Further more we screened 24 herbal abstracts and found some had the function similar to 5 alpha-reductase inhibitor, such as Jiang Huang （姜黄 ） and Xiakucao （ 夏枯草 ）. The above result proved that the non-isotope method in this study is more effective and practical, and can be used as a non-isotope model in vitro for screening 5alpha-reductase inhibitor.

关 键 词：5α-还原酶活性 筛选 非那雄胺 特异性5α-还原酶抑制剂 体外筛选模型 非同位素 NADPH 反应产物 中药提取物 二氢睾酮 酶提取物 前列腺组织 

分 类 号：R96[医药卫生—药理学]