EFFECTS OF ALCOHOL ON HUMAN OVARIAN CELL FUNCTION

酒精对人类卵巢细胞功能的影响

• 批准号: 3112386
• 负责人: JAY wimalasena WIMALASENA
• 金额: $ 15.28万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-06-01 至 1995-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3112386
• 关键词: G protein; acetaldehyde; affinity chromatography; alcohol dehydrogenase; alcoholism /alcohol abuse; amenorrhea; antiantibody; autoradiography; cellular pathology; cyclic AMP; drug interactions; enzyme linked immunosorbent assay; enzyme mechanism; female; female reproductive system disorder; fertility; gonadotropins; high performance liquid chromatography; histamine receptor; hormone binding protein; human tissue; immunological substance; immunoprecipitation; laboratory rat; menstrual cycle disorder; model design /development; monoclonal antibody; morphology; ovary; ovary disorder; pregnancy loss; protein purification; radioimmunoassay; second messengers; steroid biosynthesis; tissue /cell culture; toxicology

Alcoholism in women leads to significant aberrations in reproductive system function such as, amenorrhea, dysmenorrhea, anovulation, infertility, luteal phase defects, earlier menopause and increased rates of spontaneous abortions. Only some of these changes appear to be reversible. Recent data suggest that reproductive system dysfunction is also observed in non alcoholic women who are alcohol abusers. Even though data suggest the ethanol may exert detrimental effects on the hypothalmopituitary axis and or the ovary, the site(s) and mechanisms(s) of alcohol toxicity on the reproductive tract are poorly understood. The long term objectives of the proposed research are to understand the mechanisms of alcohol toxicity on the ovary. Previous research indicated that chronic alcoholism may produce significant ovarian dysfunction and morphological changes with consequent decreases in steroid hormone secretion, and these effects may give rise to much of the reproductive toxicity of ethanol. To understand the toxicity of alcohol on reproductive function at the molecular level it is imperative to develop suitable in vitro models. Recently a tissue culture system where human granulosa lutein (LG) cells are exposed to constant "physiological" concentrations of ethanol has been developed. In this system ethanol selectively inhibited stimulatory effects of LH on steroidogenesis. This system will be used to study the mechanism of toxicity of ethanol directly on human LG cells and human ovarian cells. Planned experiments are the following; studies of ethanol action on LH receptor number and affinity using iodo hormone binding and independent evaluation of LH receptor number with LH receptor antibodies, alcohol action on LH receptor, guanine binding protein (G protein) interactions and effects of ethanol on G protein coupling to adenylate cyclase. Since LH may act on steroidogenesis via the Ca++/inositol phosphate/diacylglycerol/protein kinase C pathway, actions of ethanol on components of this system will be measured to determine whether ethanol can modulate LH's physiologic effects via this pathway. A cultured rat LG cell model will be developed to try and duplicate the effects of alcohol on human LG cells and from these studies we hope to establish the mechanisms by which alcohol acts as a direct gonadal toxin.

女性酗酒会导致生殖系统严重畸变 功能，如闭经、痛经、不排卵、不孕、 黄体期缺陷、更年期提前和自发性妊娠率增加 堕胎。 这些变化中只有一些似乎是可逆的。 最近的 数据表明，在非生殖系统中也观察到生殖系统功能障碍。 酗酒的女性是酗酒者。 尽管数据表明 乙醇可能对下丘脑垂体轴产生有害影响 或卵巢，酒精毒性的部位和机制 人们对生殖道知之甚少。 拟议研究的长期目标是了解 酒精对卵巢的毒性机制。 先前的研究表明 长期酗酒可能会导致严重的卵巢功能障碍 形态变化导致类固醇激素减少 分泌，这些影响可能会引起许多生殖 乙醇的毒性。 了解酒精对生殖的毒性 必须在分子水平上开发合适的功能 体外模型。 最近有一个组织培养系统，其中人类颗粒 叶黄素（LG）细胞暴露于恒定的“生理”浓度 乙醇已被开发出来。 在该系统中，乙醇选择性抑制 LH 对类固醇生成的刺激作用。 该系统将用于 研究乙醇直接对人LG细胞的毒性机制 人类卵巢细胞。 计划的实验如下；研究 使用碘激素，乙醇对 LH 受体数量和亲和力的作用 与LH受体结合并独立评估LH受体数量 抗体、酒精对 LH 受体的作用、鸟嘌呤结合蛋白 (G 蛋白质）相互作用以及乙醇对 G 蛋白偶联的影响 腺苷酸环化酶。 由于 LH 可能通过 Ca++/磷酸肌醇/二酰甘油/蛋白激酶 C 途径，作用 将测量该系统组件上的乙醇以确定是否 乙醇可以通过该途径调节 LH 的生理效应。 一个有文化的 将开发大鼠 LG 细胞模型来尝试复制 酒精对人类 LG 细胞的影响，从这些研究中我们希望确定 酒精作为直接性腺毒素的机制。