[体外灌注人胎肾的类固醇生物合成(作者译)]。

M Seki, T Kimura, T Takayama, T Yanaihara, T Nakayama
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引用次数: 0

摘要

在本实验中,从两个中期流产的胎儿(胎龄21周)中获得的人胎肾在体外灌注标记的前体,包括14g -脱氢表雄酮(DHA)和14c -脱氢表雄酮硫酸(DHA- s)。灌注终止后从灌注液和组织中提取放射性代谢物,游离部分用乙醚分离,共轭部分用正丁醇分离。利用薄层色谱法对回收的放射性物质进行分离纯化。代谢产物的放射化学纯度通过再结晶达到恒定的比活性。当以DHA为前体时,在灌注液中,DHA- s、5-雄烯二醇、5-雄烯三醇被鉴定为DHA衍生的代谢物,并依次重结晶。但是,当使用DHA-S作为前体时,无法获得代谢物的放射性纯度。灌注研究的结果表明,胎儿肾脏中类固醇磺化酶活性强,类固醇16 α -羟化酶、17 β -羟基类固醇脱氢酶活性弱。
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[Biosynthesis of steroids by human fetal kidney perfused in vitro (author's transl)].

In the present experiments, human fetal kidneys obtained from two fetuses aborted at mid-term (gestational age 21 weeks) were perfused in vitro with labeled precursor including 14G-dehydroepiandrosterone (DHA) and 14C-dehydroepiandrosterone-sulfate (DHA-S). Radioactive metabolites extracted after the termination of perfusion from the perfusate and the tissue were separated the free fraction by ether and conjugate fraction by n-butanol. Furthermore, recovery of the radioactivities were separated and purified by thin layer chromatography. Radiochemical purity of the metabolites were achieved by recrystallization to constant specific activity. When DHA was used as a precursor, in the perfusate, DHA-S, delta 5-Androstenediol, delta 5-Androstenetriol were identified as the metabolites derived from DHA, and they were successively recrystallized. But, when DHA-S was used as a precursor, the radioactive purity of the metabolite was not obtained. The result of this perfusion studies demonstrate a powerful activity of steroids sulfokinase and a slight activity of steroids 16 alpha-hydroxylase, 17 beta-hydroxysteroid dehydrogenase in fetal kidney.

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