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Quantitative Evaluation and Prediction of Drug-Toxicity Due to Lipid Peroxidation

脂质过氧化引起的药物毒性的定量评估和预测

基本信息

批准号：
63571061
负责人：
HORIE Toshiharu
金额：
$ 1.41万
依托单位：
Tokyo University of Pharmacy and Life Science
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1990
项目状态：
已结题

项目摘要

Drug-induced lipid peroxidation was investigated in vitro and in vivo. 1. The method to determine glutathione was set up using HPLC which was supplied by this grant. The determination was carried out by labeling eluted glutathione with o-phthalaldehyde.2. Lipid peroxidation was evaluated by thiobarbituric acid reactive substances (TBA-RS), fluorescent substances and high molecular weight protein aggregates. Fluorescent substances formed in peroxidized microsomes of rat liver were characterized by measuring fluorescence lifetime.Fluorescent substances were found to have at least three different fluorescence lifetimes. This was the same in microsomal proteins and lipids. Microsomal enzymes decreased their activities during lipid peroxidation and membrane fluidity also decreased.3. Acetaminophen was orally administered to rats. Plasma glutamic-oxaloacetic transaminase (GOT) increased and TBA-RS in liver homogenates slightly increased. When diethylmaleate was injected intraperitoneally to rats before acetaminophen administration, fluorescent substances and high molecular weight protein aggregates were found in liver microsomes. This indicates that glutatione plays an important role in acetaminophen-induced lipid peroxidation. The same was observed in carbon tetrachloride and adriamycin. These indices to evaluate lipid peroxidation were found to be useful to detect in vivo lipid peroxidation.4. Effects of acetaminophen on isolated hepatocytes were investigated. Hepatocytes were incubated with acetaminophen. When TBA-RS was produced, cell viability decreased. Simultaneously, GOT in the medium increased and intracellular glutathione decreased. At higher concentration of acetaminophen, TBA-RS formation was depressed and cell viability was not decreased,which was probably due to the antioxidant effect of acetaminophen itself. Thus, this study showed that lipid peroxidation played an important role in acetaminophen-induced hepatotoxicity.

在体外和体内研究了药物诱导的脂质过氧化。 1. 测定谷胱甘肽的方法采用本课题提供的HPLC法建立。采用邻苯二甲醛标记洗脱的谷胱甘肽进行测定。 2．通过硫代巴比妥酸反应物质（TBA-RS）、荧光物质和高分子量蛋白质聚集体评价脂质过氧化。通过测量荧光寿命来表征大鼠肝脏过氧化微粒体中形成的荧光物质。发现荧光物质具有至少三种不同的荧光寿命。这在微粒体蛋白质和脂质中也是相同的。脂质过氧化过程中微粒体酶活性降低，膜流动性降低。3．对大鼠口服对乙酰氨基酚。血浆谷氨酸草酰乙酸转氨酶 (GOT) 升高，肝匀浆中的 TBA-RS 略有升高。在给予对乙酰氨基酚之前给大鼠腹腔注射马来酸二乙酯时，在肝微粒体中发现荧光物质和高分子量蛋白质聚集体。这表明谷胱甘肽在对乙酰氨基酚诱导的脂质过氧化中起着重要作用。在四氯化碳和阿霉素中也观察到同样的情况。这些评价脂质过氧化的指标被发现有助于检测体内脂质过氧化。4．研究了对乙酰氨基酚对分离肝细胞的影响。肝细胞与对乙酰氨基酚一起孵育。当TBA-RS产生时，细胞活力下降。同时，培养基中的GOT增加，细胞内谷胱甘肽减少。在较高浓度的对乙酰氨基酚下，TBA-RS的形成受到抑制，细胞活力并未降低，这可能是由于对乙酰氨基酚本身的抗氧化作用所致。因此，这项研究表明脂质过氧化在对乙酰氨基酚诱导的肝毒性中发挥重要作用。