Aghajanshakeri Shaghayegh, Salmanmahiny Amirhossein, Aghajanshakeri Shahin, Babaei Amirhossein, Alishahi Farhad, Babayani Erfan, Shokrzadeh Mohammad
Department of Toxicology and Pharmacology, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran.
Department of Toxicology and Pharmacology, Student Research Committee, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran.
Toxicol Mech Methods. 2023 Nov;33(9):755-765. doi: 10.1080/15376516.2023.2238069. Epub 2023 Aug 3.
Amifostine is used in chemotherapy and radiotherapy as a cytoprotective adjuvant alongside DNA-binding chemotherapeutic agents. It functions by reducing free radicals and detoxifying harmful metabolites. Methotrexate, as an antimetabolite drug has been considered for treating various cancers and autoimmune diseases. However, the cytotoxic effects of methotrexate extend beyond tumor cells to crucial organs, including the heart. This study applied the HUVEC cell line as a reference model for researching the characteristics of vascular endothelium and cardiotoxicity. The current study aimed to assess amifostine's potential cytoprotective properties against methotrexate-induced cellular damage. Cytotoxicity was measured using the MTT assay. Apoptotic rates were evaluated by Annexin V-FITC/PI staining flow cytometry. The genoprotective effect of amifostine was determined using the comet assay. Cells were exposed to various amifostine doses (10-200 μg/mL) and methotrexate (2.5 μM) in pretreatment culture condition. Methotrexate at 2.5 μM revealed cytotoxicity, apoptosis, oxidative stress and genotoxicity while highlighting amifostine's cyto/geno protective properties on HUVECs. Amifostine significantly decreased the levels of ROS and LPO while preserving the status of GSH and SOD activity. Furthermore, it inhibited genotoxicity (tail length, %DNA in tail, and tail moment) in the comet assay. Amifostine markedly attenuated methotrexate-induced apoptotic cell death (early and late apoptotic rates). These findings convey that amifostine can operate as a cytoprotectant agent.
氨磷汀在化疗和放疗中作为细胞保护佐剂与DNA结合化疗药物联合使用。它通过减少自由基和使有害代谢产物解毒发挥作用。甲氨蝶呤作为一种抗代谢药物，已被用于治疗各种癌症和自身免疫性疾病。然而，甲氨蝶呤的细胞毒性作用不仅限于肿瘤细胞，还会延伸至包括心脏在内的重要器官。本研究应用人脐静脉内皮细胞（HUVEC）系作为研究血管内皮特征和心脏毒性的参考模型。本研究旨在评估氨磷汀对甲氨蝶呤诱导的细胞损伤的潜在细胞保护特性。使用MTT法测量细胞毒性。通过Annexin V-FITC/PI染色流式细胞术评估凋亡率。使用彗星试验确定氨磷汀的基因保护作用。在预处理培养条件下，将细胞暴露于不同剂量的氨磷汀（10 - 200μg/mL）和甲氨蝶呤（2.5μM）。2.5μM的甲氨蝶呤显示出细胞毒性、凋亡、氧化应激和基因毒性，同时突出了氨磷汀对HUVECs的细胞/基因保护特性。氨磷汀显著降低了活性氧（ROS）和脂质过氧化物（LPO）水平，同时保持了谷胱甘肽（GSH）和超氧化物歧化酶（SOD）活性状态。此外，它在彗星试验中抑制了基因毒性（尾长、尾中DNA百分比和尾矩）。氨磷汀显著减轻了甲氨蝶呤诱导的凋亡细胞死亡（早期和晚期凋亡率）。这些发现表明氨磷汀可以作为一种细胞保护剂发挥作用。
