Abstract：
Cardiotoxicity is a well-established adverse effect of several drugs across multiple therapeutic indications. It is particularly prevalent following anticancer therapy. In order to evaluate the changes in cellular metabolism associated with methotrexate cardiotoxicity, we treated Wistar rats with a single high dose of methotrexate (HDMTX), and after five days, the animals were sacrificed. We then analyzed the cardiotoxicity parameters in serum like Cardiac enzymes(CK-MB, Troponin T, ALP), Inflammatory markers (TNF-α and IL-6), oxidative stress markers (NO, NOX-2), histopathology and cardiac tissue with the goal of identifying a metabolic signature of cardiotoxicity using discovery-based metabolomics. The biochemical parameters for cardiac enzymes, oxidative stress and inflammatory markers showed a significant increase in all three categories in rats treated with HDMTX. These findings were mirrored in the histopathological analysis confirming cardiotoxicity due to HDMTX. The results showed a total of 95 metabolites that were found to be significantly (p < 0.05) modulated: either up- or downregulated in the HDMTX-treated group when compared with the control group. Using integrated pathway analysis we found these metabolites were associated with many important cardiac tissue metabolic pathways, such as the malate aspartate shuttle, taurine and hypotaurine metabolism, betaine metabolism, spermidine biosynthesis, and homocysteine degradation. Among them, L-arginine, homocysteine, and betaine were significantly upregulated, suggesting their possible association with cardiac tissue injury. Overall, we provided evidence for using untargeted metabolomics to identify novel metabolites associated with HDMTX cardiac toxicity.
摘要：
心脏毒性是几种药物在多种治疗适应症中公认的不良反应。它在抗癌治疗后特别普遍。为了评估与甲氨蝶呤心脏毒性相关的细胞代谢变化，我们用单次高剂量甲氨蝶呤（HDMTX）治疗Wistar大鼠，五天后，动物被处死。然后，我们分析了血清中的心脏毒性参数，如心肌酶（CK-MB，肌钙蛋白T,ALP),炎症标志物（TNF-α和IL-6），氧化应激标志物(NO，NOX-2),组织病理学和心脏组织，目的是使用基于发现的代谢组学鉴定心脏毒性的代谢特征。心肌酶的生化参数，氧化应激和炎症标记物显示在用HDMTX治疗的大鼠中所有三个类别均显着增加。这些发现反映在组织病理学分析中，证实了由HDMTX引起的心脏毒性。结果显示,当与对照组相比时,在HDMTX处理组中发现总共95种代谢物被显著(p<0.05)调节:上调或下调。使用整合途径分析，我们发现这些代谢物与许多重要的心脏组织代谢途径有关。比如苹果酸天冬氨酸穿梭，牛磺酸和下牛磺酸代谢，甜菜碱代谢，亚精胺生物合成，和高半胱氨酸降解。其中,L-精氨酸,同型半胱氨酸,甜菜碱明显上调,提示它们可能与心脏组织损伤有关。总的来说,我们提供了使用非靶向代谢组学鉴定与HDMTX心脏毒性相关的新型代谢产物的证据.
