Effect of IC50 dose of retinol on metabolomics of RAW264.7 cells

Background: Vitamin A is one of the most multifunctional Vitamin in the human body and is involved in several basic physiological processes from embryonic development to adulthood, such as embryogenesis, vision, immunity, cell differentiation, and proliferation. The aim of current study was to identify its effects on the differential metabolites and main metabolic pathways. We also analyzed the toxicity of retinol. Methods: In this study, we conducted proton nuclear magnetic resonance (NMR) to evaluate metabolomic changes in RAW264.7 cells after treatment with retinol at a half maximal inhibitory concentration (IC 50 ). Results: Our results showed that the IC 50 dose (140 μM) of retinol affected the metabolism of RAW264.7 cells, with a total of 22 differential metabolites identified via 1 H-NMR, including amino acids, sugars, organic acids, glutathione, glycerin, and creatine. Additionally, multiple metabolic pathways were affected by retinol treatment, including amino acid biosynthesis, protein synthesis, and pyruvate metabolism. We speculate that the cytotoxicity of retinol at the IC 50 dose is attributed to mitochondrial dysfunction as a result of oxidative stress or lipid peroxidation. Conclusions: RAW264.7 cells showed significant changes in protein biosynthesis, urea cycle, arginine and proline metabolism, malate-aspartate shuttle, alanine metabolism, and cellular respiration after treatment with retinol at the IC 50 dose (140 μM), indicating that retinol affects cellular physiological functions via different metabolic pathways.