Section Environment and Health, Amsterdam Institute for Life and Environment, De Boelelaan 1085, Amsterdam, 1081 HV, The Netherlands.
Department of Gynecology and Reproductive Medicine, Karolinska University Hospital, Stockholm, SE-14186, Sweden.
Reprod Biol Endocrinol. 2024 May 22;22(1):60. doi: 10.1186/s12958-024-01233-7.
Reproduction in women is at risk due to exposure to chemicals that can disrupt the endocrine system during different windows of sensitivity throughout life. Steroid hormone levels are fundamental for the normal development and function of the human reproductive system, including the ovary. This study aims to elucidate steroidogenesis at different life-stages in human ovaries.
We have developed a sensitive and specific LC-MS/MS method for 21 important steroid hormones and measured them at different life stages: in media from cultures of human fetal ovaries collected from elective terminations of normally progressing pregnancy and in media from adult ovaries from Caesarean section patients, and follicular fluid from women undergoing infertility treatment. Statistically significant differences in steroid hormone levels and their ratios were calculated with parametric tests. Principal component analysis (PCA) was applied to explore clustering of the ovarian-derived steroidogenic profiles.
Comparison of the 21 steroid hormones revealed clear differences between the various ovarian-derived steroid profiles. Interestingly, we found biosynthesis of both canonical and "backdoor" pathway steroid hormones and corticosteroids in first and second trimester fetal and adult ovarian tissue cultures. 17α-estradiol, a less potent naturally occurring isomer of 17β-estradiol, was detected only in follicular fluid. PCA of the ovarian-derived profiles revealed clusters from: adult ovarian tissue cultures with relatively high levels of androgens; first trimester and second trimester fetal ovarian tissue cultures with relatively low estrogen levels; follicular fluid with the lowest androgens, but highest corticosteroid, progestogen and estradiol levels. Furthermore, ratios of specific steroid hormones showed higher estradiol/ testosterone and estrone/androstenedione (indicating higher CYP19A1 activity, p < 0.01) and higher 17-hydroxyprogesterone/progesterone and dehydroepiandrosterone /androstenedione (indicating higher CYP17A1 activity, p < 0.01) in fetal compared to adult ovarian tissue cultures.
Human ovaries demonstrate de novo synthesis of non-canonical and "backdoor" pathway steroid hormones and corticosteroids. Elucidating the steroid profiles in human ovaries improves our understanding of physiological, life-stage dependent, steroidogenic capacity of ovaries and will inform mechanistic studies to identify endocrine disrupting chemicals that affect female reproduction.
由于暴露于化学物质，女性的生殖功能受到威胁，这些化学物质在生命不同的敏感窗口可能会破坏内分泌系统。甾体激素水平对人类生殖系统的正常发育和功能至关重要，包括卵巢。本研究旨在阐明人类卵巢在不同生命阶段的甾体生成。
我们开发了一种灵敏且特异的 LC-MS/MS 方法，可检测 21 种重要的甾体激素，并在不同的生命阶段进行测量：从正常进行的妊娠终止中收集的人胎儿卵巢培养物的培养基中，以及从剖腹产患者的成人卵巢中，以及接受不孕治疗的女性的卵泡液中。使用参数检验计算甾体激素水平及其比值的统计学差异。应用主成分分析（PCA）探索卵巢源性甾体生成谱的聚类。
对 21 种甾体激素的比较揭示了各种卵巢源性甾体谱之间存在明显差异。有趣的是，我们发现第一和第二孕期胎儿和成人卵巢组织培养物中均存在经典和“旁路”途径甾体激素和皮质类固醇的生物合成。17α-雌二醇，一种比 17β-雌二醇活性低的天然存在的异构体，仅在卵泡液中检测到。卵巢源性谱的 PCA 显示出聚类：成人卵巢组织培养物中雄激素水平相对较高；第一和第二孕期胎儿卵巢组织培养物中雌激素水平相对较低；卵泡液中雄激素水平最低，但皮质类固醇、孕激素和雌二醇水平最高。此外，特定甾体激素的比值显示出更高的雌二醇/睾酮和雌酮/雄烯二酮（表明 CYP19A1 活性更高，p<0.01）和更高的 17-羟孕酮/孕酮和脱氢表雄酮/雄烯二酮（表明 CYP17A1 活性更高，p<0.01）在胎儿与成人卵巢组织培养物中。
人类卵巢表现出非经典和“旁路”途径甾体激素和皮质类固醇的从头合成。阐明人类卵巢的甾体生成谱可提高我们对卵巢生理、生命阶段依赖的甾体生成能力的理解，并为识别影响女性生殖的内分泌干扰化学物质的机制研究提供信息。
