DNA, PROTEINS & CARBON TETRACHLORIDE HEPATOTOXICITY

该项目的长期目标是获得有关如何 自由基引起的化学物质，后来遗传物质和/或其 通过研究肝毒素CC 14对肝DNA的影响， 和不同的核（n）蛋白质。 观察到的影响将是 与已知的肝脏明显不同的反应相关， 从不同的动物物种或品系到CC 14作用（坏死， 肝硬化、癌症）（从现在起在文中称为“肝脏反应 CC14”）。 本项目的具体目标是：（a）核实是否有 14 CC 14与DNA或与DNA的共价结合（CB）之间的相关性 不同的n-蛋白组分和已知的“对CC 14的肝脏反应”， 不同的动物种类或品系。 （a）开展结构研究 当CC 13自由基与DNA碱基反应时形成的加合物 和氨基酸。 （c）验证是否存在相关性， CC 1促进“体内”或“体外”脂质过氧化（LP）的能力 （肝切片、微粒体和细胞核）和已知的“肝对 不同动物种属或品系的CC 14”。 （d）核实是否 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”与DNA或正- proteins. e）研究CC 13诱导的DNA碱基和氨基酸的改变， 不能归因于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， 不同的N蛋白级分（例如，组蛋白、酸性蛋白等）并 确定CB的范围。 B）尝试识别 HPLC法测定碱基或核苷和氨基酸的改变 (UV/14 C/荧光、电测检测）和/或通过GLC/MS在 不同的实验条件。（e）确定CC 14是否促进 通过乙烷或丙二醛或4测定的核LP 羟基壬烯醛生产。 计划对健康的主要影响： 理解自由基诱导的DNA和N蛋白的改变 及其在急性和长期化学诱导的 肝损伤 结果可能会给一个深入了解的作用机制， 弱致突变性或无致突变性的肝致癌物，如CC 14。

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.