Thurston L M, Norgate D P, Jonas K C, Chandras C, Kloosterboer H J, Cooke B A, Michael A E
Department of Biochemistry and Molecular Biology, Royal Free and University College Medical School, University College London, Rowland Hill Street, London NW3 2PF, UK.
Reproduction. 2002 Dec;124(6):801-12. doi: 10.1530/rep.0.1240801.
In the ovary, cortisol-cortisone interconversion is catalysed by isoforms of 11beta-hydroxysteroid dehydrogenase (11betaHSD). The objective of this study was to establish whether human follicular fluid (hFF), obtained after controlled ovarian hyperstimulation, contains paracrine modulators of 11betaHSD activity. Of 274 hFF samples tested for effects in rat kidney homogenates, 206 hFF samples significantly inhibited NADP(+)-dependent oxidation of cortisol within 1 h (by 11-67% of control 11betaHSD activity), whereas 42 hFF samples significantly stimulated 11betaHSD activity (16-210% increase relative to control). Although charcoal-stripping of hFF prevented the inhibition and potentiated the stimulation of NADP(+)-dependent cortisol oxidation in a renal homogenate, effects of individual hFF samples on NADP(+)-dependent cortisol oxidation were independent of intrafollicular progesterone concentrations. Hydrophilic fractions of hFF samples, isolated by C18 column chromatography, stimulated both the NADP(+)-dependent oxidation of cortisol (by 55+/-5%, n=98) and the NADPH-dependent reduction of cortisone (by 86+/-22%, n= 5). In contrast, the hydrophobic fractions of hFF (eluted at 65-85% methanol) inhibited both NADP(+)-dependent 11beta-dehydrogenase and NADPH-dependent 11-ketosteroid reductase activities (by 63+/-2% and 74+/-4%, respectively). None of the C18 column fractions of 50 hFF samples had any significant effect on NAD(+)-dependent 11beta-dehydrogenase activities. The hydrophobic inhibitors of NADP(H)-dependent cortisol-cortisone metabolism did not co-elute with several candidate compounds (prostaglandins E(2) and F(2alpha), cortisol, cortisone, oestradiol, testosterone, progesterone, pregnenolone or cholesterol). Hence, hFF aspirated from women undergoing controlled ovarian hyperstimulation for assisted conception contains both hydrophilic stimuli and hydrophobic inhibitors of glucocorticoid metabolism which appear to be selective for the NADP(H)-dependent, type 1 isoform of 11betaHSD.
在卵巢中，皮质醇与可的松的相互转化由11β-羟基类固醇脱氢酶（11βHSD）的同工型催化。本研究的目的是确定在控制性卵巢过度刺激后获得的人卵泡液（hFF）是否含有11βHSD活性的旁分泌调节因子。在对274份hFF样本进行大鼠肾匀浆效应测试中，206份hFF样本在1小时内显著抑制了NADP(+)-依赖性皮质醇氧化（为对照11βHSD活性的11%-67%），而42份hFF样本显著刺激了11βHSD活性（相对于对照增加16%-210%）。尽管hFF的活性炭处理可防止肾匀浆中对NADP(+)-依赖性皮质醇氧化的抑制并增强刺激作用，但单个hFF样本对NADP(+)-依赖性皮质醇氧化的影响与卵泡内孕酮浓度无关。通过C18柱色谱分离的hFF样本的亲水性部分，既刺激了NADP(+)-依赖性皮质醇氧化（增加55±5%，n = 98），也刺激了NADPH依赖性可的松还原（增加86±22%，n = 5）。相反，hFF的疏水性部分（在65%-85%甲醇中洗脱）抑制了NADP(+)-依赖性11β-脱氢酶和NADPH依赖性11-酮类固醇还原酶活性（分别为63±2%和74±4%）。50份hFF样本的C18柱馏分对NAD(+)-依赖性11β-脱氢酶活性均无显著影响。NADP(H)-依赖性皮质醇-可的松代谢的疏水性抑制剂未与几种候选化合物（前列腺素E2和F2α、皮质醇、可的松、雌二醇、睾酮、孕酮、孕烯醇酮或胆固醇）共洗脱。因此，从接受控制性卵巢过度刺激以进行辅助受孕的女性中抽取的hFF含有糖皮质激素代谢的亲水性刺激物和疏水性抑制剂，它们似乎对NADP(H)-依赖性11βHSD 1型同工型具有选择性。
