PAH ACTION ON OVARIAN STEROIDOGENESIS IN VITRO

描述：（根据申请人的描述改编） 该建议的假设是多环芳烃（PAH） 是内分泌干扰物， 妇女生殖健康的直接影响 通过细胞变性损害卵巢卵泡健康 （闭锁）/程序性细胞死亡（凋亡）和通过改变类固醇 合成能力。 有人提出，这种环境异生物质 通过诱导细胞凋亡的发生率增加来发挥这些作用， 卵泡颗粒细胞（GC）;并影响一个或多个 在生物合成途径中的几个位点：在P450侧链裂解， 3b-羟基类固醇脱氢，P45017，20裂解酶裂解，和/或在 芳构化水平。 TCDD和另一种AH受体（AHR）激动剂， 亲和力较低的β-萘酚酮（BNF）将在类器官中给药 组织块培养成年Holtzman大鼠完整卵泡的研究 还涉及人颗粒细胞的原代细胞培养物， 接受体外受精手术的妇女;以及对发病率的影响 细胞凋亡和类固醇生成的影响。 具体来说，调查人员 将表征AHR激动剂对（A）以下检测的影响： 应用DNA片段免疫荧光定位原位凋亡 （B）在细胞化学上定位参与和 用生化/放射性方法分析大鼠卵泡膜的转化， 线粒体胆固醇转化为双烯醇酮（P5）;（C）P5转化为 孕酮（P4）在滑面内质网;（D）微粒体 P5裂解成脱氢表雄酮;和（E）P5的芳构化 雄烯二酮对雌酮在大鼠卵泡和人颗粒细胞中的作用。 的 研究人员将把形态学变化和 生物合成，其特征为毒素诱导的卵泡死亡。

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.