Synthesis of free and sulphoconjugated 16-androstene steroids by the Leydig cells of the mature domestic boar
Synthesis of free and sulphoconjugated 16-androstene steroids by the Leydig cells of the mature domestic boar
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DOI：
10.1016/j.jsbmb.2005.02.017
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发表时间：
2005-07-01
影响因子：
4.1
通讯作者：
Renaud, R
中科院分区：
文献类型：
--
作者：
Sinclair, PA;Squires, EJ;Renaud, R
这项研究考察了硫代偶联作用在成熟猪的间质细胞中16-雄烯类固醇的生物合成中的作用，因为类固醇硫代偶合物的形成可以减少这些类固醇在脂肪组织中积累的水平。从成年公猪睾丸中分离纯化睾丸间质细胞，分别与孕烯醇酮或不同的16-雄烯类固醇孵育10min、1、4、8h后，用固相萃取-溶剂法回收硫代偶联类固醇。用高效液相色谱法分析了未结合和硫代结合的甾体化合物的谱带。硫代偶联组分中存在的类固醇被提纯，衍生化为O-甲酮肟/三甲基硅基醚(MO-TMS)，然后用气相色谱-质谱仪(GC-MS)进行鉴定。孕烯醇酮、雄二烯醇、雄二烯酮和5α-雄烯酮与孕烯醇酮孵育产生的主要代谢产物为3β-雄烯醇。硫代偶联的16-雄烯类化合物包括5-α-雄烯酮、3-β-雄烯醇和3-α-雄烯醇。在孵育4小时或更长时间后，每个16-雄烯类固醇总量的大约70%以硫代偶联的形式存在。硫代偶联的5α-雄烯酮的大量存在表明，这种类固醇可能从3-酮转化为3-烯醇形式，然后再进行硫代偶联。这些发现强调了需要考虑16-雄烯类固醇的硫代偶联的影响及其在造成野猪污染中的作用。(C)2005爱思唯尔有限公司。保留所有权利。
This study examined the involvement of sulphoconjugation in the biosynthesis of the 16-androstene steroids in Leydig cells of the mature boar, since the formation of steroid sulphoconjugates can reduce the levels of these steroids that accumulate in fatty tissue. Leydig cells were purified from testes of mature male pigs and incubated with pregnenolone, or various individual 16-androstene steroids for 10 min, 1, 4 and 8 h. Sulphoconjugated steroids were recovered by solid-phase extraction followed by solvolysis. Profiles of unconjugated and sulphoconjugated steroids were analysed by HPLC. Steroids present in the sulphoconjugated fractions were purified, derivatised as O-methoxime/trimethylsilyl ethers (MO-TMS), and subsequently identified using gas chromatography-mass spectrometry (GC-MS). The principal metabolite produced from incubations with pregnenolone, androstadienol, androstadienone and 5 alpha-androstenone was 3 beta-androstenol. 16-Androstene steroids that were sulphoconjugated included 5 alpha-androstenone, 3 beta-androstenol and 3 alpha-androstenol. Approximately 70% of the total amount of each 16-androstene steroid was in its sulphoconjugated form after incubations for 4 h or more. The finding that sulphoconjugated 5 alpha-androstenone was present in large amounts suggests that this steroid may be converted from a 3-keto to a 3-enol form which is subsequently sulphoconjugated. These findings emphasise the need to consider the impact of sulphoconjugation of the 16-androstene steroids and their role in contributing to boar taint. (c) 2005 Elsevier Ltd. All rights reserved.