Pharmacokinetics and Hepatotoxicity due to the oxidative stress

为了验证活性氧在药物肝毒性中的作用，检测药物代谢过程中产生的活性氧是非常重要的。用化学发光法研究了药物诱导的大鼠肝脏灌流、离体肝细胞和线粒体的氧化应激反应。1.用醋氨酚溶液灌流大鼠肝匀浆，发现其产生超微弱化学发光，并在肝匀浆中形成高分子量蛋白质聚集体，还原型谷胱甘肽减少。用萘普生溶液灌流的大鼠肝脏中氧化型谷胱甘肽略有增加。2.对乙酰氨基酚在离体肝细胞中产生超微弱化学发光。萘普生还可使肝细胞悬液产生化学发光，并增加氧化型谷胱甘肽占总谷胱甘肽的比例。依他尼酸由于氧化代谢而诱导肝细胞中的脂质过氧化。3.在线粒体中检测到阿霉素的超弱化学发光，并且发现化学发光的起源是单线态氧和受激羰基。用荧光-高效液相色谱法检测了阿霉素给药小鼠心脏匀浆中磷脂酰胆碱过氧化氢的含量。

In order to verify the contribution of the active oxygens to the drug-induced hepatotoxicity, it is very important to detect the active oxygens generated during the drug metabolism. Chemiluminescence produced from the active oxygens, glutathione, fluorescent substances, high molecular weight protein aggregates and thiobarbituric acid reactive substances were used to study the drug-induced oxidative stress in perfused rat liver, isolated rat hepatocyte and mitochondria. 1. The ultra weak chemiluminescence was detected in the homogenates of rat liver perfused with acetaminophen solution.The high molecular weight protein aggregates were foromed in the liver perfused with a cetaminophen and the reduced glutathione decreased in the liver. The oxidized glutathione increased slightly in the rat liver perfused with naproxen solution. 2. The ultra weak chemiluminescence was detected in the isolated hepatocytes with acetaminophen. Naproxen also produced the chemiluminescence in the hepatocyte suspension and increased the ratio of oxidized glutathione to the total glutathione. Ethacrynic acid induced lipid peroxidation in the hepatocytes due to the oxidative metabolism. 3. The ultra weak chemiluminescence was detected in the mitochondria with doxorubicin and the origins of the chemiluminescence were found to be singlet oxygen and excited carbonyls. Phosphatidylcholine hydroperoxide was detected in the heart homogenates of doxorubicin administered mouse using the chemiluminescence-HPLCsystem.