Al-Nasser Moonerah M, Al-Saeedi Mashael J, Alhowaiti Saltana A, Shinwari Zakia, Alhamlan Fatimah S, Alothaid Hani, Alkahtani Saad, Al-Qahtani Ahmed A
Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia.
Department of Infection and Immunity, Research Centre, King Faisal Specialist Hospital & Research Centre, Riyadh, Saudi Arabia.
J Inflamm Res. 2024 Nov 2;17:8085-8098. doi: 10.2147/JIR.S482503. eCollection 2024.
Methotrexate (MTX) is a widely used anti-metabolite drug in cancer therapy, but its efficacy is often hindered by reactive oxygen species (ROS)-induced cellular toxicity. Resveratrol, a natural polyphenol, possesses antioxidant and anticancer properties. Therefore, this in vitro study aimed to investigate the synergistic anti-proliferative and anti-inflammatory effects of MTX and resveratrol in human THP-1 cells.
THP-1 cells were differentiated into macrophage-like cells using phorbol 12-myristate 13-acetate. In vitro experiments assessed the impact of various concentrations of MTX and resveratrol on cell viability and proliferation using the MTT assay. Concentration-effect curves were generated to elucidate their relationship. Gene expression was analyzed by RT-qPCR, while chemokine expression was measured via ELISA. Phagocytic and migratory activities were also evaluated.
Differentiated THP-1 cells were treated with MTX and resveratrol and stimulated with dimethyl sulfoxide (DMSO) and lipopolysaccharide (LPS) as inflammatory stimuli. The combination of MTX and resveratrol exhibited an anti-proliferative effect in THP-1 cells (p = 0.03). The concentration-effect curve revealed IC50 values of 49.15 µg at 24 hours (R = 0.8236) and 2.029e-005 µg at 48 hours (R = 0.97) for MTX, and 20.17 µg at 48 hours (R = 1.000) and 55.38 µg at 96 hours (R = 0.9666) for resveratrol. Co-treatment with MTX and resveratrol significantly reduced mRNA and chemokine expression of CCL2, CCL3, CCL4, CCL5, and CXCL10 (p < 0.05). Moreover, decreased phagocytic and migratory activities were confirmed by phagocytosis and migration assays (p < 0.05).
The combination of MTX and resveratrol effectively attenuated pro-inflammatory activity in THP-1 cells, as evidenced by the downregulation of mRNA and chemokine expression. These findings suggest that the synergistic effects of MTX and resveratrol hold promise for enhancing cancer therapeutics.
甲氨蝶呤（MTX）是癌症治疗中广泛使用的抗代谢药物，但其疗效常受活性氧（ROS）诱导的细胞毒性阻碍。白藜芦醇是一种天然多酚，具有抗氧化和抗癌特性。因此，本体外研究旨在探讨MTX与白藜芦醇在人THP-1细胞中的协同抗增殖和抗炎作用。
使用佛波醇12-肉豆蔻酸酯13-乙酸酯将THP-1细胞分化为巨噬细胞样细胞。体外实验通过MTT法评估不同浓度的MTX和白藜芦醇对细胞活力和增殖的影响。绘制浓度-效应曲线以阐明它们之间的关系。通过RT-qPCR分析基因表达，同时通过ELISA测定趋化因子表达。还评估了吞噬和迁移活性。
用MTX和白藜芦醇处理分化的THP-1细胞，并用二甲基亚砜（DMSO）和脂多糖（LPS）作为炎症刺激物进行刺激。MTX与白藜芦醇的组合在THP-1细胞中表现出抗增殖作用（p = 0.03）。浓度-效应曲线显示，MTX在24小时时的IC50值为49.15μg（R = 0.8236），在48小时时为2.029e-005μg（R = 0.97）；白藜芦醇在48小时时的IC50值为20.17μg（R = 1.000），在96小时时为55.38μg（R = 0.9666）。MTX与白藜芦醇联合处理显著降低了CCL2、CCL3、CCL4、CCL5和CXCL10的mRNA和趋化因子表达（p < 0.05）。此外，吞噬和迁移实验证实吞噬和迁移活性降低（p < 0.05）。
MTX与白藜芦醇的组合有效减弱了THP-1细胞中的促炎活性，mRNA和趋化因子表达下调证明了这一点。这些发现表明MTX与白藜芦醇的协同作用有望增强癌症治疗效果。
