POTENTIAL FETOPROTECTANTS FROM ETOH INDUCED STRESS

免受乙醇引起的应激的潜在胎儿保护剂

• 批准号: 2865453
• 负责人: Linda S LaGrange
• 金额: $ 9.1万
• 依托单位: NEW MEXICO HIGHLANDS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-03 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2865453
• 关键词: S adenosylmethionine; acetylcysteine; alcoholic beverage consumption; antioxidants; blood chemistry; body weight; cytoprotection; cytotoxicity; disease /disorder model; enzyme activity; ethanol; fetal alcohol syndrome; flavones; free radicals; gestational age; glutathione; hepatotoxin; laboratory rat; lipid peroxides; neuroprotectants; neurotoxins; oxidative stress; prenatal stress; quercetin; tocopherols

The ingestion of EtOH can lead to the creation of a variety of free radical species. If EtOH consumption is heavy and chronic, organs within the body begin to deteriorate. Some of the organ damage sustained as a consequence of EtOH consumption might be caused by free radical generation and subsequent lipid peroxidation (LPO). It is now theorized that the pervasive and toxic effects of LPO may contribute to the expression of fetal alcohol syndrome (FAS). Silymarin (SY), a plant-derived flavone, has been identified as an effective antioxidant and is currently marketed in Europe wider the trade name Legalon(R) as a hepatoprotective medication. We have conducted several animal studies of the hepatoprotective and fetoprotective effects of SY against EtOH-induced enzyme activity in maternal and fetal brain and liver tissue. We now want to test the antioxidant effects of SY and four other antioxidant flavonoids; quercetin, myricetin, (+)-catechin, and apigenin on EtOH-exposed fetal rat brain and liver tissue. Testing of the additional bioflavonoid compounds is predicated on the results of studies in which it was demonstrated that these flavonoids also inhibited EtOH-induced free radical production. For purposes of comparison of antioxidant efficacy, N-acetylcysteine (NAC) and S- adenosylmethione (SAM) will also be tested. It has been demonstrated in previous in vitro studies that the incubation of fetal rat hepatocyte (FRH) mitochondria in EtOH increased indications of oxidative stress. It was further established that the treatment of the FRH mitochondria with NAC or SAM before and during the 24-hr exposure to EtOH prevented the decrease in glutathione (GSH), restored cell replication, and moderated malondialdehyde (MDA) production. We will extend this in vitro model to an in vivo model to test the general hypothesis that one or all of the flavonoids will be as effective as NAC or SAM at normalizing Et0H-induced oxidative stress in fetal brain and liver tissue. Dams will be intubated with EtOH six times at 12-hr. intervals over days 12, 13, and 14 of gestation following the procedures described in Henderson, Devi, Perez, & Schencker (1995). The dependent measures used to assess outcome will be: fetal viability, fetal weight, liver histology, fetal brain and liver tissue glutathione levels, alpha tocopherol levels, MDA levels, and GGTP activity. In addition, to continue with current studies in our lab, we will establish parallel groups in which rat pups will be raised for subsequent behavioral testing.

摄入ETOH会导致产生各种自由基物种。如果ETOH消耗量很重且慢性，则体内内脏开始恶化。由于ETOH消耗而导致的某些器官损伤可能是由于自由基产生和随后的脂质过氧化（LPO）引起的。现在认为，LPO的普遍和毒性作用可能有助于胎儿酒精综合征（FAS）的表达。植物来源的植物衍生的黄酮（SY）已被确定为有效的抗氧化剂，目前在欧洲销售的商品名称Legallon（R）作为一种肝保护药物。我们已经进行了几项动物研究，该研究对孕产妇和胎儿脑和肝组织中ETOH诱导的酶活性的肝保护作用和胎儿保护作用。现在，我们想测试SY和其他四种抗氧化剂类黄酮的抗氧化作用。槲皮素，米他汀，（+） - 儿茶素和阿apegenin在eTOH暴露的胎儿大鼠脑和肝组织上。额外的生物黄酮化合物的测试基于研究结果，在研究结果表明这些类黄酮还抑制了EtOH诱导的自由基生产。为了比较抗氧化功效，还将测试N-乙酰半胱氨酸（NAC）和S-腺苷甲硫代（SAM）。在先前的体外研究中已经证明，在ETOH中孵育胎儿大鼠肝细胞（FRH）线粒体的孵育增加了氧化应激的指示。进一步确定的是，在24小时接触ETOH之前和期间，通过NAC或SAM处理FRH线粒体可以防止谷胱甘肽（GSH）减少（GSH），恢复的细胞复制和调节的丙二醛（MDA）产生。我们将将这种体外模型扩展到体内模型，以检验一个普遍的假设，即一种或全部的类黄酮将与NAC或SAM一样有效，可以使胎儿脑和肝组织中ET0H诱导的氧化应激归一化。大坝将在12小时时用ETOH插管六次。妊娠第12、13和14天的间隔按照亨德森，德维，佩雷斯和舒恩克（1995）所述的程序。用于评估结果的依赖措施将是：胎儿活力，胎儿体重，肝组织学，胎儿脑和肝组织谷胱甘肽水平，α生育酚水平，MDA水平和GGTP活性。 此外，为了继续在我们的实验室进行当前的研究，我们将建立平行组，其中将饲养大鼠幼崽以进行随后的行为测试。