DNA, PROTEINS & CARBON TETRACHLORIDE HEPATOTOXICITY

DNA、蛋白质

• 批准号: 3253885
• 负责人: JOSE A CASTRO
• 金额: $ 9.34万
• 依托单位: LELOIR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3253885
• 关键词: DNA; adduct; aminoacid; arachidonate; carbon tetrachloride poisoning; chemical binding; chemical carcinogenesis; covalent bond; free radical oxygen; gas chromatography mass spectrometry; hamsters; hepatotoxin; high performance liquid chromatography; laboratory mouse; laboratory rat; lipids; liver cirrhosis; liver neoplasms; liver toxic disorder; male; necrosis; nucleoproteins; nucleosides; peroxidation; protein purification; scintillation counter; toxin metabolism

The long term objective of the project are to gain knowledge baout how free radical causing chemicals later genetic material and/or its expression, by studying the effects of the hepatotoxin CC14 on liver DNA and different nuclear (n) proteins. The observed effects would be correlated with the already known markedly different response of livers from different animal species or strains to CC14 action (necrosis, cirrhosis, cancer) (from now on in text referred as the "liver response to CC14"). The present project specific aims are: a) To verify whether there is a correlation between the covalent binding (CB) of 14CC14 to DNA or to different n-protein fractions and the known "liver response to CC14" of different animal species or strains. a) To initiate structural studies of the adducts formed when the CC13 free radicals react with DNA bases and amino acids. c) To verify whether there is a correlation between CC1 ability to promote lipid peroxidation (LP) "in vivo" or "in vitro" (liver slices, micorsomes and nuclei) and the known "liver response to CC14" of different animal species or strains. d) To verify whether arachidonic acid degradation products produced during LP (e.g., 4 hydroxynonenal) are able to CB "in vivo" or "in vitro" to DNA or n- proteins. e) To study CC13 induced alterations in DNA bases and amino acids not attributable to either CB or LP (e.g., cross links formation, breakdown). To achieve these goals we plan: a) To islate highly purified DNA and different n-protein fractions (e.g., histones, acidic, etc.) and to establish the extent of CB to them. b) To attempt identification of altered bases or nucleosides and amino acids by HPLC (UV/14C/fluoroscence.electrometric detection) and/or by GLC/MS under different experimental conditions. e) To establish whether CC14 promotes nuclear LP as determined by ethanepenthane or malondialdehyde or 4 hydroxynonenal production. Major Health Implications of The Project: To achieve a better understanding of free radical induced alterations in DNA and n-proteins and their potential role in acute and long term chemically n=induced liver damage. Results might give an insight to mechanisms of action of liver carcinogens of weak or null mutagenic nature as is CC14.

该项目的长期目标是获得有关如何 产生自由基的化学物质后来的遗传物质和/或其 通过研究肝毒素 CC14 对肝脏 DNA 的影响 和不同的核（n）蛋白。 观察到的效果是 与已知的肝脏显着不同的反应相关 来自不同动物物种或品系的 CC14 作用（坏死、 肝硬化、癌症）（从现在开始在文本中称为“肝脏反应 至 CC14”）。 本项目的具体目标是： a) 验证是否存在 14CC14 与 DNA 或与 DNA 的共价结合 (CB) 之间的相关性 不同的 n 蛋白部分和已知的“CC14 肝脏反应” 不同的动物种类或品系。 a) 启动结构研究 CC13 自由基与 DNA 碱基反应时形成的加合物 和氨基酸。 c) 验证之间是否存在相关性 CC1“体内”或“体外”促进脂质过氧化 (LP) 的能力 （肝切片、微粒体和细胞核）和已知的“肝脏反应” 不同动物物种或品系的CC14"。 d) 验证是否 LP 过程中产生的花生四烯酸降解产物（例如 4 羟基壬烯醛）能够“体内”或“体外”CB 到 DNA 或 n- 蛋白质。 e) 研究 CC13 诱导的 DNA 碱基和氨基的改变 不可归因于 CB 或 LP 的酸（例如，交联形成， 分解）。 为了实现这些目标，我们计划： a) 分离高度纯化的 DNA 并 不同的 n 蛋白组分（例如组蛋白、酸性蛋白等）并 确定他们的 CB 范围。 b) 尝试识别 通过 HPLC 改变碱基或核苷和氨基酸 (UV/14C/荧光.静电检测)和/或通过GLC/MS在 不同的实验条件。 e) 确定 CC14 是否促进 核LP由乙烷或丙二醛测定或4 羟基壬烯醛的生产。 该项目的主要健康影响：实现更好的 了解自由基诱导的 DNA 和 n 蛋白改变 及其在急性和长期化学 n=诱导中的潜在作用 肝脏损伤。 结果可能有助于了解作用机制 弱或无诱变性的肝癌致癌物，如 CC14。