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.

女性酗酒导致生殖系统严重异常 闭经、痛经、无排卵、不孕不育、 黄体期缺陷、更年期提前和自发性增加 堕胎。这些变化中似乎只有一部分是可逆的。近期 数据表明，生殖系统功能障碍也在非 酗酒的女性酗酒者。尽管数据表明 乙醇可能对下丘脑垂体轴产生不利影响 或卵巢，酒精中毒的部位(S)和机制(S) 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 慢性酒精中毒可能会导致严重的卵巢功能障碍 伴随着类固醇激素下降的形态变化 分泌，而这些影响可能会引起大量的生殖 乙醇的毒性。了解酒精对生殖系统的毒性 在分子水平上发挥作用，势在必行 体外模型。最近，一种组织培养系统，其中人颗粒 叶黄素(LG)细胞暴露于恒定的生理浓度的 乙醇已经开发出来了。在这个体系中，乙醇被选择性地抑制 促黄体生成素对类固醇合成的促进作用。这个系统将被用来 乙醇对人LG细胞直接毒性作用机制的研究 人类卵巢细胞。计划中的实验如下；研究 用碘激素研究乙醇对黄体生成素受体数目和亲和力的影响 黄体生成素受体数目与黄体生成素受体的结合和独立评估 抗体、酒精对黄体生成素受体、鸟嘌呤结合蛋白的作用(G 蛋白质)相互作用及乙醇对G蛋白偶联的影响 腺苷环化酶。由于黄体生成素可能通过 Ca++/肌醇磷酸/甘油二酯/蛋白激酶C途径的作用 将测量乙醇对该系统组件的影响，以确定是否 乙醇可通过这一途径调节黄体生成素的生理效应。一个有教养的人 将建立大鼠LG细胞模型以尝试复制 酒精对人LG细胞的影响，我们希望从这些研究中建立 酒精作为直接性腺毒素的作用机制。