Immunomodulatory Effects of Bacillus natto Glycopeptides on RAW264.7 Macrophages
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Abstract:
The immunomodulatory effects of Bacillus natto glycopeptides (BNGP) on normal and lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages were investigated in the present study. After treatment with different concentrations of BNGP or combinations of LPS and BNGP in different concentrations, the proliferation of macrophages was detected by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, the phagocytic activity was detected by neutral red phagocytosis experiments, and the content of nitric oxide (NO) in the cell culture supernatants was detected by the Griess reaction. The production of cytokines (IL-1β, TNF-α) and pro-inflammatory mediator (PGE2) in culture supernatants were measured using an enzyme-linked immunosorbent assay (ELISA), and the expression levels of iNOS and COX-2 were detected using western blotting. The results showed that at concentrations of 62.5–500 μg/mL, BNGP enhanced the metabolic activity of macrophage RAW264.7 in the normal steady state, increased the secretions of NO, IL-1β, TNF-α, and PGE2, and improved the expression levels of iNOS and COX-2; at high concentrations (>125 μg/mL), it enhanced the phagocytic capacity of macrophages. For the LPS-stimulated macrophages, BNGP inhibited the secretions of IL-1β, TNF-α, and PGE2 at concentrations of 62.5–500 μg/mL, inhibited the production of NO and the expression of iNOS at concentrations of 500 μg/mL, and downregulated the expression of COX-2 at high concentrations (>125 μg/mL).
