Combination of Methotrexate and Resveratrol Reduces Pro-Inflammatory Chemokines in Human THP-1 Cells

Moonerah M Al-Nasser; Mashael J Al-Saeedi; Saltana A Alhowaiti; Zakia Shinwari; Fatimah S Alhamlan; Hani Alothaid; Saad Alkahtani; Ahmed A Al-Qahtani

doi:10.2147/JIR.S482503

Combination of Methotrexate and Resveratrol Reduces Pro-Inflammatory Chemokines in Human THP-1 Cells

J Inflamm Res. 2024 Nov 2:17:8085-8098. doi: 10.2147/JIR.S482503. eCollection 2024.

Authors

Moonerah M Al-Nasser¹, Mashael J Al-Saeedi², Saltana A Alhowaiti², Zakia Shinwari³, Fatimah S Alhamlan^{2

4}, Hani Alothaid⁵, Saad Alkahtani¹, Ahmed A Al-Qahtani^{2

4}

Affiliations

¹ Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia.
² Department of Infection and Immunity, Research Centre, King Faisal Specialist Hospital & Research Centre, Riyadh, Saudi Arabia.
³ Therapeutics & Biomarker Discovery for Clinical Applications, Stem Cell & Tissue Re-Engineering Program, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia.
⁴ Department of Microbiology and Immunology, College of Medicine, Alfaisal University, Riyadh, Saudi Arabia.
⁵ Department of Basic Sciences, Faculty of Applied Medical Sciences, Al-Baha University, Al-Baha, Saudi Arabia.

Abstract

Introduction: 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.

Methods: 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.

Results: 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).

Conclusion: 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.

Keywords: THP-1; anti-inflammatory; immune cells; immunomodulatory; methotrexate; resveratrol.