(1)H-NMR Metabolomics Analysis of the Effects of Sulfated Polysaccharides from Masson Pine Pollen in RAW264.7 Macrophage Cells

Many polysaccharides have been shown to be bioactive, with the addition of sulfate often enhancing or altering this bioactivity. In previous studies, masson pine pollen polysaccharides, to include a sulfate derivative, have been shown to promote macrophage proliferation similarly to LPS. However, the exact metabolic mechanisms promoting this proliferation remain unclear. In this study, RAW264.7 macrophage cells were treated with a purified masson pine pollen polysaccharide (PPM60-D), a sulfate derivative (SPPM60-D), or LPS. Proliferation levels at a variety of concentrations were examined using MTT assay, with optimal concentration used when performing metabolomic analysis via (1)H nuclear magnetic resonance ((1)H-NMR). This process resulted in the identification of thirty-five intracellular metabolites. Subsequent multivariate statistical analysis showed that both LPS and SPPM60-D promote RAW264.7 proliferation by promoting aerobic respiration processes and reducing processes associated with glycolysis. While some insight was gained regarding the mechanistic differences between SPPM60-D and LPS, the specific mechanisms governing the effect of SPPM60 on RAW264.7 cells will require further elucidation. These findings show that both LPS and SPPM60-D effectively promote RAW264.7 proliferation and may have beneficial uses in maintaining cellular vitality or inhibiting cancer.