Role of metabolites of cyclophosphamide in cardiotoxicity.
Role of metabolites of cyclophosphamide in cardiotoxicity.
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DOI：
10.1186/s13104-017-2726-2
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发表时间：
2017-08-14
影响因子：
1.8
通讯作者：
Kawano Y
中科院分区：
文献类型：
--
作者：
Kurauchi K;Nishikawa T;Miyahara E;Okamoto Y;Kawano Y
环磷酰胺（CY）的剂量限制性毒性作用是心脏毒性。心肌损伤的发病机制知之甚少，也没有既定的预防手段。在以前的研究中，我们认为，对于CY诱导的心脏毒性，而丙烯醛是关键的毒性代谢产物，羧乙基磷酰胺氮芥（CEPM）是保护性的。我们试图验证丙烯醛是心脏毒性的主要原因，并调查是否醛脱氢酶（ALDH），这是与更大的CEPM生产，参与心脏毒性的保护作用。我们还评估了N-乙酰半胱氨酸（NAC）的保护作用，NAC是一种具有抗氧化活性的氨基酸，也是一种已知的丙烯醛清除剂。将H9 c2细胞暴露于CY代谢物HCY（4-羟基-环磷酰胺）、丙烯醛或CEPM。通过MTT法、乳酸脱氢酶（LDH）释放和活性氧（ROS）产生来评价细胞毒性程度。我们还研究了NAC如何改变CY代谢物的心肌细胞保护作用。为了定量丙烯醛水平，我们使用高效液相色谱法测量培养物上清液。我们测量了暴露于HCY或丙烯醛后的ALDH活性，以及NAC预处理后的ALDH活性。H9 c2细胞暴露于CEPM不会引起细胞毒性。然而，HCY和丙烯醛可诱导ROS水平升高和心肌细胞毒性。在细胞培养中，HCY被代谢为丙烯醛。HCY或丙烯醛暴露后，ALDH活性降低。用NAC处理降低丙烯醛浓度。增加的ROS生成和降低的ALDH活性证实了CY代谢物HCY和丙烯醛与心脏毒性密切相关。通过抑制ROS的产生，增加ALDH活性和减少丙烯醛的存在，NAC有可能防止CY诱导的心脏毒性。
The dose-limiting toxic effect of cyclophosphamide (CY) is cardiotoxicity. The pathogenesis of myocardial damage is poorly understood, and there is no established means of prevention. In previous studies, we suggested that for CY-induced cardiotoxicity, whereas acrolein is the key toxic metabolite, carboxyethylphosphoramide mustard (CEPM) is protective. We sought to verify that acrolein is the main cause of cardiotoxicity and to investigate whether aldehyde dehydrogenase (ALDH), which is associated with greater CEPM production, is involved in the protective effect for cardiotoxicity. We also evaluated the protective effect of N-acetylcysteine (NAC), an amino acid with antioxidant activity and a known acrolein scavenger. H9c2 cells were exposed to CY metabolites HCY (4-hydroxy-cyclophosphamide), acrolein or CEPM. The degree of cytotoxicity was evaluated by MTT assay, lactate dehydrogenase (LDH) release, and the production of reactive oxygen species (ROS). We also investigated how the myocardial cellular protective effects of CY metabolites were modified by NAC. To quantify acrolein levels, we measured the culture supernatants using high performance liquid chromatography. We measured ALDH activity after exposure to HCY or acrolein and the same with pre-treatment with NAC. Exposure of H9c2 cells to CEPM did not cause cytotoxicity. Increased ROS levels and myocardial cytotoxicity, however, were induced by HCY and acrolein. In cell cultures, HCY was metabolized to acrolein. Less ALDH activity was observed after exposure to HCY or acrolein. Treatment with NAC reduced acrolein concentrations. Increased ROS generation and decreased ALDH activity confirmed that CY metabolites HCY and acrolein are strongly implicated in cardiotoxicity. By inhibiting ROS generation, increasing ALDH activity and decreasing the presence of acrolein, NAC has the potential to prevent CY-induced cardiotoxicity.