Methotrexate modulates folate phenotype and inflammatory profile in EA.hy 926 cells

Area of Science:

• Endothelial Cell Biology
• Pharmacology
• Molecular Biology

Background:

• Endothelial cells (EA.hy 926) under low folate conditions exhibit a pro-atherosclerotic phenotype.
• Methotrexate (MTX), an antifolate drug, is used clinically but its effects on endothelial cells require further investigation.

Purpose of the Study:

• To investigate the impact of methotrexate on EA.hy 926 cells under varying folate conditions.
• To analyze the transcriptomic and biochemical changes induced by MTX in endothelial cells.

Main Methods:

• EA.hy 926 cells were cultured in normal (Hi) and low (Lo) folate media and treated with MTX.
• Cell proliferation and metabolic activity were assessed.
• Transcript profiling was performed using microarrays.
• Quantitative RT-PCR and ELISA were used to validate gene expression changes.

Main Results:

• MTX inhibited cell proliferation but not metabolic activity in EA.hy 926 cells.
• MTX treatment led to folate derivative depletion and altered proportions in Hi cells.
• Transcriptomic analysis revealed significant upregulation of inflammation-related genes (C3, IL-8) and downregulation of cell division genes.
• C3 and IL-8 upregulation was confirmed by RT-PCR and ELISA.

Conclusions:

• Methotrexate modifies the inflammatory potential of EA.hy 926 cells.
• MTX treatment can induce gene products that promote or maintain comorbid pathologies, potentially alongside therapeutic benefits.