Thomas F J, Thomas M J, Tetsuka M, Mason J I, Hillier S G
Department of Obstetrics and Gynaecology, University of Edinburgh, UK.
Clin Endocrinol (Oxf). 1998 Apr;48(4):509-13. doi: 10.1046/j.1365-2265.1998.00457.x.
The aims of this study were to determine the type and level of 11 beta-hydroxysteroid dehydrogenase (11 beta HSD) in human granulosa-leutein cells (GLE) shortly before ovulation and to correlate activity with the outcome of treatment in patients undergoing in vitro fertilization and embryo transfer (IVF/ET).
GLC from 32 patients undergoing IVF/ET were tested for type and level of 11 beta HSD activity in relation to treatment outcome.
Periovulatory follicles were aspirated by ultrasound guided transvaginal puncture following a standard controlled ovarian stimulation protocol, approximately 36 h after administering an ovulation-inducing dose of human chorionic gonadotrophin (HCG). GLC were separated from follicular fluid by density-gradient centrifugation and taken for measurement of 11 beta HSD activity in vitro; oocytes were used for IVF/ET.
Interconversion of cortisol (F) and cortisone (E), and dexamethasone (D) and 11-dehydrodexamethasone (DHD) was measured in standardized assays comprising incubation of GLC with 3H-labelled substrate, with separation of substrate and product by thin-layer radiochromatography.
Conversion of F to E varied from 10.5 to 30.9% while that of E to F was between 2.4 and 44.6%. In the GLC of 25 patients in whom both activities were measured, dehydrogenase (F to E) activity predominated in 13 and reductase (E to F) in 12. By contrast, D (substrate for 11 beta HSD2 but not 11 beta HSD1) showed less than 1% metabolism in this system while DHD (substrate for 11 beta HSD1 and 11 beta HSD2) was converted significantly (65.6-90.5%) to D in the four patients tested. There was no significant difference in the interconversion of F and E between patients who became pregnant and those who did not.
The dehydrogenase and oxoreductase reactions catalysed by 11 beta HSD both occur in granulosa-lutein cells at the time of follicular rupture, probably due to 11 beta HSD1. A lack of measurable conversion of dexamethasone to 11-dehydrodexamethasone suggests that dehydrogenation due to 11 beta HSD2 is low or absent. Neither type nor level of 11 beta HSD activity measured under the present assay conditions correlates with IVF outcome.
本研究旨在确定排卵前不久人颗粒黄体细胞（GLE）中11β-羟基类固醇脱氢酶（11βHSD）的类型和水平，并将其活性与接受体外受精和胚胎移植（IVF/ET）患者的治疗结果相关联。
对32例接受IVF/ET患者的GLC进行检测，以确定11βHSD活性的类型和水平与治疗结果的关系。
按照标准的控制性卵巢刺激方案，在给予人绒毛膜促性腺激素（HCG）诱导排卵剂量约36小时后，通过超声引导经阴道穿刺抽吸排卵周围卵泡。通过密度梯度离心从卵泡液中分离出GLC，并用于体外测量11βHSD活性；卵母细胞用于IVF/ET。
在标准化检测中测量皮质醇（F）和可的松（E）以及地塞米松（D）和11-脱氢地塞米松（DHD）的相互转化，检测包括将GLC与3H标记的底物孵育，通过薄层放射色谱法分离底物和产物。
F向E的转化率在10.5%至30.9%之间，而E向F的转化率在2.4%至44.6%之间。在25例同时测量了两种活性的患者的GLC中，脱氢酶（F向E）活性在13例中占主导，还原酶（E向F）活性在12例中占主导。相比之下，D（11βHSD2而非11βHSD1的底物）在该系统中的代谢率低于1%，而DHD（11βHSD1和11βHSD2的底物）在4例检测患者中显著转化为D（65.6 - 90.5%）。怀孕患者和未怀孕患者之间F和E的相互转化没有显著差异。
11βHSD催化的脱氢酶和氧化还原酶反应在卵泡破裂时的颗粒黄体细胞中均会发生，可能是由于11βHSD1。地塞米松向11-脱氢地塞米松缺乏可测量的转化表明，11βHSD2引起的脱氢作用较低或不存在。在本检测条件下测量的11βHSD活性的类型和水平均与IVF结果无关。
