Sultan C, Terraza A, Devillier C, Carilla E, Briley M, Loire C, Descomps B
J Steroid Biochem. 1984 Jan;20(1):515-9. doi: 10.1016/0022-4731(84)90264-4.
We previously suggested [Steroids 33, (1979) 3; Steroids 37, (1981) 6] that cultured genital skin fibroblasts should prove useful for screening of potential antiandrogens in human and living target cells. "Serenoa repens" lipidic extract (S.R.E.) was recently reported (Br. J. Pharmacol., in press) to inhibit androgen action in animals. The present investigation was designed to study the antiandrogenicity of this compound in human cells: we therefore analyzed the effects of S.R.E. on the intracellular conversion of testosterone (T) to 5 alpha-reduced derivatives, and we investigated interaction of S.R.E. with the intracellular androgen-receptor complex. Since the chemical structure of the active component of S.R.E. is still unknown, results are expressed in U/ml (one unit is defined as the amount of S.R.E. required to inhibit 50% of the specific binding (IC50) of [3H]1881 to rat prostate cytosol). S.R.E. at different dilutions (5.7 to 28.6 U/ml) is added to culture media containing [3H]T or [3H]dihydrotestosterone (DHT) and incubated at 37 degrees C with cultured fibroblasts. 28.6 U/ml S.R.E. significantly alters the formation of DHT and strongly inhibits 3 ketosteroid reductase mediated conversion of DHT to 5 alpha-androstane-3 alpha, 17 beta-diol, characterized radiochemically by thin-layer chromatography. S.R.E. is a good competitor for the whole cell androgen receptor: 7.1 U/ml S.R.E. gives 50% inhibition of the binding of 2 X 10(-9) M [3H]DHT to its receptor. Competitive binding assays after cell fractionation indicate that S.R.E. is less potent in nuclear than in cytosol receptors. Sucrose gradient centrifugation of the radioactive cell lysate of fibroblasts demonstrates that 28.6 U/ml S.R.E. abolishes 70% of the 3.6 S receptor-complex radioactive peak. The present studies show that S.R.E. inhibits 5 alpha-reductase, 3-ketosteroid reductase and receptor binding of androgens in cultured human foreskin fibroblasts. As the search for the ideal antiandrogen continues, S.R.E. appears to be a new type of antiandrogenic compound as therapeutics for the treatment of benign prostatic hypertrophy, hirsutism and so forth.
我们之前曾提出[《类固醇》33卷，（1979年）第3期；《类固醇》37卷，（1981年）第6期]，培养的生殖器皮肤成纤维细胞应可用于在人类及活体靶细胞中筛选潜在的抗雄激素药物。最近有报道（《英国药理学期刊》，即将发表）称，“锯叶棕”脂质提取物（S.R.E.）可抑制动物体内的雄激素作用。本研究旨在探讨该化合物在人类细胞中的抗雄激素性：因此，我们分析了S.R.E.对睾酮（T）向5α-还原衍生物的细胞内转化的影响，并研究了S.R.E.与细胞内雄激素受体复合物的相互作用。由于S.R.E.活性成分的化学结构仍不清楚，结果以U/ml表示（一个单位定义为抑制[3H]1881与大鼠前列腺胞质溶胶特异性结合（IC50）的50%所需的S.R.E.量）。将不同稀释度（5.7至28.6 U/ml）的S.R.E.添加到含有[3H]T或[3H]双氢睾酮（DHT）的培养基中，并与培养的成纤维细胞在37℃下孵育。28.6 U/ml的S.R.E.显著改变了DHT的形成，并强烈抑制了3-酮类固醇还原酶介导的DHT向5α-雄甾烷-3α，17β-二醇的转化，通过薄层色谱进行放射化学表征。S.R.E.是全细胞雄激素受体的良好竞争者：7.1 U/ml的S.R.E.可使2×10^(-9) M [3H]DHT与其受体的结合受到50%的抑制。细胞分级分离后的竞争性结合试验表明，S.R.E.在核受体中的效力低于胞质溶胶受体。对成纤维细胞放射性细胞裂解物进行蔗糖梯度离心表明，28.6 U/ml的S.R.E.使3.6 S受体复合物放射性峰消失了70%。目前的研究表明，S.R.E.可抑制培养的人类包皮成纤维细胞中5α-还原酶、3-酮类固醇还原酶以及雄激素的受体结合。随着对理想抗雄激素药物的探索仍在继续，S.R.E.似乎是一种新型的抗雄激素化合物，可用于治疗良性前列腺增生、多毛症等。
