Faculty of Pharmacy and Pharmaceutical Sciences, 2142 J Katz Group-Rexall Centre for Pharmacy and Health Research, University of Alberta, Edmonton, AB, T6G 2E1, Canada.
Cardiovasc Toxicol. 2018 Jun;18(3):268-283. doi: 10.1007/s12012-017-9437-8.
Several studies have demonstrated the role of cytochrome P450 (CYP) and its associated arachidonic acid (AA) metabolites in the anthracyclines-induced cardiac toxicity. However, the ability of daunorubicin (DNR) to induce cardiotoxicity through the modulation of CYP and its associated AA metabolites has not been investigated yet. Therefore, we hypothesized that DNR-induced cardiotoxicity is mediated through the induction of cardiotoxic hydroxyeicosatetraenoic acids and/or the inhibition of cardioprotctive epoxyeicosatrienoic acids (EETs). To test our hypothesis, Sprague-Dawley rats were treated with DNR (5 mg/kg i.p.) for 24 h, whereas human ventricular cardiomyocytes RL-14 cells were exposed to DNR in the presence and absence of 4-[[trans-4-[[(tricyclo[3.3.1.13,7]dec-1-ylamino)carbonyl]amino]cyclohexyl]oxy]-benzoic acid (tAUCB), a soluble epoxide hydrolase (sEH) inhibitor. Thereafter, real-time PCR, Western blot analysis and liquid chromatography-electron spray ionization mass spectroscopy were used to determine the level of gene expression, protein expression and AA metabolites, respectively. Our results showed that DNR-induced cardiotoxicity in vivo and in vitro as evidenced by the induction of hypertrophic and fibrotic markers. Moreover, the DNR-induced cardiotoxicity was associated with a dramatic increase in the formation of cardiac DHET/EET metabolites both in vivo and in RL-14 cells suggesting a sEH enzyme dependent mechanism. Interestingly, inhibition of sEH using tAUCB, a selective sEH inhibitor, significantly protects against DNR-induced cardiotoxicity. Mechanistically, the protective effect tAUCB was mediated through the induction of P50 nuclear factor-κB and the inhibition of phosphorylated p38. In conclusion, our study provides the first evidence that DNR induces cardiotoxicity through a sEH-mediated EETs degradation-dependent mechanism.
几项研究已经证明细胞色素 P450（CYP）及其相关的花生四烯酸（AA）代谢物在蒽环类抗生素诱导的心脏毒性中的作用。然而，尚未研究柔红霉素（DNR）通过调节 CYP 及其相关的 AA 代谢物诱导心脏毒性的能力。因此，我们假设 DNR 诱导的心脏毒性是通过诱导心脏毒性羟二十碳四烯酸和/或抑制心脏保护环氧二十碳三烯酸（EETs）来介导的。为了验证我们的假设，用 DNR（5mg/kg ip）处理 Sprague-Dawley 大鼠 24h，而 RL-14 人心肌细胞则在存在和不存在 4-[[反式-4-[[（三环[3.3.1.13,7]癸-1-基氨基）羰基]氨基]环己基]氧基] - 苯甲酸（tAUCB）的情况下暴露于 DNR 中，tAUCB 是一种可溶性环氧化物水解酶（sEH）抑制剂。此后，实时 PCR、Western blot 分析和液相色谱-电子喷雾电离质谱分别用于确定基因表达、蛋白表达和 AA 代谢物的水平。我们的结果表明，DNR 诱导体内和体外的心脏毒性，表现为诱导肥大和纤维化标志物。此外，DNR 诱导的心脏毒性与心脏 DHET/EET 代谢物形成的急剧增加有关，这表明存在 sEH 酶依赖性机制。有趣的是，使用 tAUCB（一种选择性 sEH 抑制剂）抑制 sEH 可显著防止 DNR 诱导的心脏毒性。机制上，tAUCB 的保护作用是通过诱导 P50 核因子-κB 和抑制磷酸化 p38 来介导的。总之，我们的研究首次提供了证据，表明 DNR 通过 sEH 介导的 EETs 降解依赖性机制诱导心脏毒性。
