PAH ACTION ON OVARIAN STEROIDOGENESIS IN VITRO

PAH 对体外卵巢类固醇生成的作用

• 批准号: 2518711
• 负责人: REINHOLD Josef HUTZ
• 金额: $ 13.65万
• 依托单位: UNIVERSITY OF WISCONSIN MILWAUKEE
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2518711
• 关键词: aromatic hydrocarbon receptor; carbopolycyclic compound; cell death; dioxins; endocrine pharmacology; environmental toxicology; female; granulosa cell; hormone regulation /control mechanism; human tissue; immunocytochemistry; immunofluorescence technique; in situ hybridization; laboratory rat; reproductive system pharmacology; steroid biosynthesis

DESCRIPTION: (adapted from the applicant's description) The general hypothesis of this proposal is that polycyclic aromatic hydrocarbons (PAH) are endocrine-disrupting chemicals that exert their untoward cellular effects on female reproductive health by a direct action at the level of the ovary via impairing ovarian follicular health through cellular degeneration (atresia)/programmed cell death (apoptosis) and by altering steroid synthetic capability. It is proposed that such environmental xenobiotics exert these effects by inducing an increased incidence of apoptosis in follicular granulosa cells (GC); and affect steroidogenesis at one or more of several loci in the biosynthetic pathway: at P450 side-chain cleavage, 3b-hydroxysteroid dehydrogenation, P45017,20 lyase cleavage, and/or at the level of aromatization. TCDD and another AH receptor (AHR) agonist with lesser affinity, beta-naphthoflavone (BNF), will be administered in organoid tissue culture to intact ovarian follicles from adult cycling Holtzman rats and also to primary cell cultures of human granulosa cells retrieved from women undergoing in vitro fertilization procedures; and effects on incidence of apoptosis and steroidogenesis evaluated. Specifically, the investigators will characterize the effects of AHR agonists on (A) the detection of apoptosis in situ using immunofluorescence localization of DNA fragmented internucleosomally; (B) localize cytochemically the enzymes involved in and analyze biochemically/radiometrically the conversion of rat thecal mitochondrial cholesterol to pregnenolone (P5); (C) the conversion of P5 to progesterone (P4) in smooth endoplasmic reticulum; (D) the microsomal cleavage of P5 to dehydroepiandrosterone; and (E) the aromatization of androstenedione to estrone in rat follicles and human granulosa cells. The investigators will correlate morphologic changes and alterations in biosynthesis that characterize toxin-induced demise of ovarian follicles.

描述：(改编自申请者的描述)将军 这一提议的假设是多环芳烃(PAH) 是扰乱内分泌的化学物质，对细胞产生不利影响 在妇女一级采取直接行动对女性生殖健康的影响 卵巢通过细胞变性损害卵泡健康 (闭锁)/程序性细胞死亡(细胞凋亡)和改变类固醇 综合能力。有人提出，这种环境异生物质 通过诱导更多的细胞凋亡来发挥这些作用 卵泡颗粒细胞(GC)；并影响一个或多个类固醇的生成 在生物合成途径中的几个基因座：在P450侧链断裂时， 3B-羟基类固醇脱氢，P45017，20裂解酶裂解，和/或 芳构化程度。TCDD和另一种AH受体(AHR)激动剂 亲和力较弱的β-萘黄酮(BNF)将以有机类药物的形式给药 成年周期Holtzman大鼠完整卵泡的组织培养 也用于原代培养的人颗粒细胞，从 接受体外受精手术的妇女及其对发病率的影响 对细胞凋亡和类固醇激素生成的影响进行了评估。具体地说，调查人员 将表征AHR激动剂对(A)检测 DNA片段化免疫荧光定位原位细胞凋亡 核内；(B)细胞化学定位参与和 大鼠卵泡膜转化率的生化/放射计量学分析 线粒体胆固醇转化为孕烯醇酮(P5)；(C)P5转化为 滑面内质网中的孕酮(P4)；(D)微体 P5裂解成脱氢表雄酮；和(E)芳构化 雄烯二酮对大鼠卵泡和人颗粒细胞雌酮的影响。这个 研究人员将把形态变化和血管的变化联系起来 毒素诱导的卵泡死亡的生物合成特征。