Metabolomic analysis reveals the influence of IC50 vitamin D3 on RAW264.7 cells based on 1H NMR and UPLC-MS/MS

Background:As a lipid-soluble vitamin necessary for normal human physiology, vitamin D is mostly used in fortified foods, medicines and adjuvant treatment of diseases. However, taken in high doses, vitamin D can be toxic.

Methods:We treated RAW264.7 cells with a semi-inhibitory concentration (IC₅₀ ) of vitamin D₃ . The metabolic changes in the treated cells were analyzed by ¹ H nuclear magnetic resonance (NMR) and ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).

Results:After treatment of RAW264.7 cells with an IC₅₀ dose of 55 μm vitamin D₃ , tunor necrosis factor-α levels decreased significantly and remarkable metabolic differences were also observed, with 12 types of metabolites were identified by ¹ H NMR and 87 identified by UPLC-MS/MS. Moreover, the metabolism of amino acids, sugars, lipids and other metabolic pathways were also affected.

Conclusion:Although vitamin D₃ is an indispensable nutrient in the body, excessive exposure has negative effects on cells and their metabolism. The present study will assist further analyses of the mechanism underlying vitamin D₃ toxicity. © 2022 Society of Chemical Industry.

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