Wildlife Research Center, Kyoto University, Kanrin, Inuyama, Aichi 484-8506, Japan.
Faculty of Science, Okayama University of Science, 1-1, Ridai-Cho, Kita-Ku, Okayama 700-0005, Japan.
Gen Comp Endocrinol. 2023 Jul 1;338:114277. doi: 10.1016/j.ygcen.2023.114277. Epub 2023 Mar 23.
We developed a microplate enzyme immunoassay (EIA) to measure dehydroepiandrosterone sulfate (DHEAS) in the blood, urine, and feces of Japanese macaques and evaluated the relationship between serum DHEAS and excreta DHEAS. Our DHEAS EIA using heterological DHEA derivatives conjugated with enzyme was highly sensitive, and linearities and recoveries for all matrices of Japanese macaques were reliable. For the biological evaluation of the metabolism of DHEAS in Japanese macaques, dissolved DHEAS was injected into the subjects, and consecutively collected serum, urine, and fecal samples were analyzed. The peaks of serum DHEAS were observed 6 h after the administration, while those of urine and feces were observed after 24 h. The fluctuation of those in urine and feces were significantly correlated with serum DHEAS levels. In addition, we measured pregnanediol-glucuronide (PdG), and estrone-conjugate (EC) in urine and fecal samples to investigate the effects of administrated DHEAS on these progesterone and estrogen metabolites. The peak of PdG was observed 24 h after administration, then declined sharply. The concentration of EC increased 1 week after administration in two out of three subjects. Our results suggest that measuring urinary and fecal DHEAS with our EIA provides a non-invasive alternative to assessing DHEAS levels in the serum of Japanese macaques.
我们开发了一种微孔板酶免疫分析(EIA)来测量日本猕猴血液、尿液和粪便中的硫酸脱氢表雄酮(DHEAS),并评估了血清 DHEAS 与粪便 DHEAS 之间的关系。我们的使用异源性 DHEA 衍生物与酶偶联的 DHEAS EIA 具有高度的灵敏度,并且所有日本猕猴基质的线性和回收率都是可靠的。为了评估 DHEAS 在日本猕猴体内的代谢情况,我们将溶解的 DHEAS 注入实验对象体内,然后连续采集血清、尿液和粪便样本进行分析。给药后 6 小时观察到血清 DHEAS 的峰值,而尿液和粪便中的峰值则在 24 小时后观察到。尿液和粪便中的波动与血清 DHEAS 水平呈显著相关性。此外,我们还测量了尿液和粪便中的孕烷二醇葡萄糖醛酸(PdG)和雌酮共轭物(EC),以研究给予的 DHEAS 对这些孕激素和雌激素代谢物的影响。PdG 的峰值在给药后 24 小时观察到,然后急剧下降。在三个实验对象中的两个中,给药后 1 周内 EC 的浓度增加。我们的结果表明,使用我们的 EIA 测量尿液和粪便中的 DHEAS 为评估日本猕猴血清中的 DHEAS 水平提供了一种非侵入性的替代方法。