Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Alberta, Canada.
Drug Metab Dispos. 2012 Nov;40(11):2126-35. doi: 10.1124/dmd.112.046631. Epub 2012 Aug 6.
Doxorubicin [(DOX) Adriamycin] is an effective anticancer agent whose major limiting side effect is cardiotoxicity. This cardiotoxicity is predicted only by the cumulative dose of DOX where the clinical situation involves chronic drug administration. Therefore, we investigate the effect of chronic DOX cardiotoxicity on expression of the cardiac cytochrome P450 (P450) enzymes and arachidonic acid (AA) metabolism in male Sprague-Dawley (SD) rats. The chronic toxicity was induced by multiple intraperitoneal injections for a cumulative dose of 15 mg/kg divided into six injections within 2 weeks. After 14 days of the last injection, the heart, liver, and kidney were harvested, and the expression of different genes was determined by real-time polymerase chain reaction. In addition, microsomal protein from the heart was prepared and incubated with AA. Thereafter, different AA metabolites were analyzed by liquid chromatography-electrospray ionization-mass spectrometry. The chronic DOX cardiotoxicity significantly induced gene expression of hypertrophic markers, apoptotic markers, CYP2E1, CYP4A3, CYP4F1, CYP4F5, and soluble epoxide hydrolase (sEH) enzyme, which was accompanied by an increase in the activity of P450 ω-hydroxylases and sEH. In addition, both the sEH inhibitor, trans-4-[4-(3-adamantan-1-yl-ureido)-cyclohexyloxy]-benzoic acid, and the ω-hydroxylase inhibitor, N-hydroxy-N'-(4-butyl-2-methylphenyl)-formamidine (HET0016), significantly prevented the DOX-mediated induction of the hypertrophic markers in the cardiac-derived H9c2 cells, which further confirms the role of these enzymes in DOX cardiotoxicity. Furthermore, gene expression of P450 and sEH was altered in an organ-specific manner. As a result, the chronic DOX administration leads to an imbalance between P450-mediated cardiotoxic and cardioprotective pathways. Therefore, P450 ω-hydroxylases and sEH might be considered as novel targets to prevent and/or treat DOX cardiotoxicity.
多柔比星（DOX）阿霉素是一种有效的抗癌药物，其主要的限制副作用是心脏毒性。这种心脏毒性只能通过 DOX 的累积剂量来预测，而临床情况涉及慢性药物给药。因此，我们研究了慢性 DOX 心脏毒性对雄性 Sprague-Dawley（SD）大鼠心脏细胞色素 P450（P450）酶和花生四烯酸（AA）代谢的表达的影响。通过多次腹腔内注射诱导慢性毒性，累积剂量为 15mg/kg，在 2 周内分为 6 次注射。最后一次注射后 14 天，采集心脏、肝脏和肾脏，通过实时聚合酶链反应测定不同基因的表达。此外，还从心脏中提取微粒体蛋白，并与 AA 孵育。然后，通过液相色谱-电喷雾电离质谱分析不同的 AA 代谢物。慢性 DOX 心脏毒性显著诱导了肥大标志物、凋亡标志物、CYP2E1、CYP4A3、CYP4F1、CYP4F5 和可溶性环氧化物水解酶（sEH）酶的基因表达，同时增加了 P450 ω-羟化酶和 sEH 的活性。此外，sEH 抑制剂 trans-4-[4-(3-金刚烷-1-基-脲基)-环己氧基]-苯甲酸和 ω-羟化酶抑制剂 N-羟基-N'-(4-丁基-2-甲基苯基)-甲脒（HET0016）均能显著预防 DOX 诱导的心脏源性 H9c2 细胞肥大标志物的诱导，这进一步证实了这些酶在 DOX 心脏毒性中的作用。此外，P450 和 sEH 的基因表达呈器官特异性改变。因此，慢性 DOX 给药导致 P450 介导的心脏毒性和心脏保护途径之间的失衡。因此，P450 ω-羟化酶和 sEH 可能被认为是预防和/或治疗 DOX 心脏毒性的新靶点。
