Wasser S K, Monfort S L, Southers J, Wildt D E
Department of Obstetrics and Gynecology, University of Washington, Seattle 98105.
J Reprod Fertil. 1994 May;101(1):213-20. doi: 10.1530/jrf.0.1010213.
Two unanaesthetized female yellow baboons (Papio cynocephalus cynocephalus) were infused (i.v.) with [3H]oestradiol and two with [3H]progesterone, early in the follicular phases of their cycles. One month later, the two females infused with [3H]oestradiol were simultaneously infused with [14C]progesterone and [3H]dehydroepiandrosterone. All urine and faeces were collected for 96 h after infusion. The proportion of steroid excreted in faeces (versus urine) was 10.0% for oestradiol and 40% for progesterone. Peak excretion in urine occurred 4.5 h after infusion. Peak excretion in faeces occurred an average of 36.4 h after infusion, with remarkable consistency between steroids. Eighty per cent of faecal oestradiol and progesterone metabolites were excreted as free (rather than conjugated) steroids. Simply boiling (20 min) the dried faecal sample in 90% ethanol proved to be the most rapid and efficient means of extracting these steroid metabolites. High pressure liquid chromatography and immunoreactivity studies revealed that oestradiol was excreted in faeces as oestradiol (36%), oestrone (44%) and a conjugated metabolite that co-eluted with oestrone sulfate (20%). Progesterone was excreted as eight different free forms, only a minor portion of which was progesterone, and what appeared to be a conjugated metabolite that co-eluted with pregnanediol-glucuronide (20%). The free progesterone metabolites were identified by gas-chromatography-mass-spectrometry as epimers of 5-pregnane-3-diol and 5-pregnane-3-ol-one. These data suggest that currently available immunoassays for free oestradiol and oestrone should adequately characterize faecal oestrogen profiles in baboons. However, high variability in crossreactivities of various progesterone antisera to progesterone metabolites in baboons makes antiserum selection a more serious concern in attempts to quantify faecal progestogen dynamics.(ABSTRACT TRUNCATED AT 250 WORDS)
在卵泡期早期,对两只未麻醉的雌性黄狒狒(豚尾狒狒)静脉注射[3H]雌二醇,对另外两只静脉注射[3H]孕酮。一个月后,对注射了[3H]雌二醇的两只雌性同时静脉注射[14C]孕酮和[3H]脱氢表雄酮。注射后96小时收集所有尿液和粪便。雌二醇经粪便排泄的比例(相对于尿液)为10.0%,孕酮为40%。尿液中排泄高峰出现在注射后4.5小时。粪便中排泄高峰平均出现在注射后36.4小时,不同类固醇之间具有显著的一致性。80%的粪便雌二醇和孕酮代谢物以游离(而非结合)类固醇的形式排泄。将干燥的粪便样本在90%乙醇中简单煮沸(20分钟)被证明是提取这些类固醇代谢物的最快速、有效的方法。高压液相色谱和免疫反应性研究表明,雌二醇以雌二醇(36%)、雌酮(44%)和一种与硫酸雌酮共洗脱的结合代谢物(20%)的形式经粪便排泄。孕酮以八种不同的游离形式排泄,其中只有一小部分是孕酮,还有一种似乎是与孕二醇 - 葡萄糖醛酸共洗脱的结合代谢物(20%)。游离孕酮代谢物通过气相色谱 - 质谱法鉴定为5 - 孕烷 - 3 - 二醇和5 - 孕烷 - 3 - 醇 - 酮的差向异构体。这些数据表明,目前可用的游离雌二醇和雌酮免疫测定法应能充分表征狒狒粪便中的雌激素谱。然而,各种孕酮抗血清对狒狒孕酮代谢物的交叉反应性存在高度变异性,这使得在试图量化粪便孕激素动态时,抗血清的选择成为一个更严重的问题。(摘要截短至250字)