Kaymak Güllü
Simav Vocational School of Health Services, Kütahya Health Sciences University, Kütahya, Türkiye.
J Med Food. 2025 Jul;28(7):657-667. doi: 10.1089/jmf.2024.0261. Epub 2025 Apr 24.
Methotrexate (MTX), a folate antimetabolite, is a cytotoxic drug known to cause cytotoxicity associated with free oxygen radicals. This study investigated the effect of vitamins D3 and K2 on MTX-induced liver cell injury using the zebrafish liver cell line (CRL-2643). Observed effects, levels of antioxidant enzymes, lipid peroxidation marker, and total antioxidant/oxidant status were evaluated by spectrophotometric methods. The mRNA expressions of p53, Bax, and Bcl-2 were measured using RT-PCR. In addition, acridine orange/ethidium bromide staining was performed to analyze the apoptosis status of the cells. The IC value of MTX at 48 h was calculated as 442 g/mL with an MTT assay. The doses of D3&K2 are determined based on the recommended dose to be taken daily. As a result, this study suggests that MTX treatment induced oxidative damage on the liver cell, as assessed by increased reactive oxygen levels, lipid peroxidation, and decreased glutathione levels. In addition, while MTX increased the expression of the p53 and proapoptotic marker Bax, it decreased the expression of the anti-apoptotic factor Bcl-2. In conclusion, D3&K2 treatment protects against MTX-induced liver cell toxicity. It is thought that they can be used as a potential agent in clinical applications with MTX in treatment.
甲氨蝶呤（MTX）是一种叶酸抗代谢物，是一种已知会导致与游离氧自由基相关的细胞毒性的细胞毒性药物。本研究使用斑马鱼肝细胞系（CRL-2643）研究了维生素D3和K2对MTX诱导的肝细胞损伤的影响。通过分光光度法评估观察到的效应、抗氧化酶水平、脂质过氧化标志物以及总抗氧化/氧化状态。使用RT-PCR测量p53、Bax和Bcl-2的mRNA表达。此外，进行吖啶橙/溴化乙锭染色以分析细胞的凋亡状态。通过MTT法计算MTX在48小时时的IC值为442μg/mL。D3和K2的剂量根据每日推荐服用剂量确定。结果，本研究表明，通过活性氧水平升高、脂质过氧化和谷胱甘肽水平降低评估，MTX处理对肝细胞诱导了氧化损伤。此外，虽然MTX增加了p53和促凋亡标志物Bax的表达，但它降低了抗凋亡因子Bcl-2的表达。总之，D3和K2处理可防止MTX诱导的肝细胞毒性。人们认为它们可以作为MTX治疗临床应用中的潜在药物。
