Effect of Bacillus natto Glycopeptide on Nuclear Translocation of Nuclear Factor-κB in RAW264.7 Macrophages
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Abstract:
The effect of Bacillus natto glycopeptide (BNGP) on the nuclear translocation of nuclear factor (NF)-κB p65 into the nucleus of RAW264.7 macrophages was studied using immunofluorescence; its effect on NF-κB p65 expression in the cytoplasm and nucleus was detected by western blot, while its effect on fluorescein isothiocyanate-lipopolysaccharide (LPS-FITC) binding to cell receptors was determined by flow cytometry. The results showed that the percentage of the cells with NF-κB p65 nuclear translocation in 62.5~500 μg/mL BNGP groups was significantly higher than that in the control group. High-dose BNGP (500 μg/mL) significantly reduced the percentage of cells with NF-κB p65 nuclear translocation induced by LPS and no significant changes were found between the low dose (≤ 250 μg/mL) BNGP and LPS groups. When macrophages were treated with 62.5~500 μg/mL BNGP alone, NF-κB p65 expression level increased in the nucleus and decreased in the cytoplasm, suggesting that BNGP promoted the translocation of NF-κB p65 from cytoplasm to nucleus. When macrophages were treated with 62.5~500 μg/mL BNGP and LPS, the expression level of NF-κB p65 increased in the cytoplasm and decreased in the nucleus, suggesting that BNGP inhibited the LPS-induced nuclear translocation of NF-κB p65 from the cytoplasm to the nucleus in RAW264.7 cells. BNGP also reduced the amount of LPS-FITC bound to macrophages at doses ranging from 62.5 to 500 μg/mL, where 500 μg/mL BNGP showed a significant effect, indicating that BNGP interferes with the binding of LPS-FITC to macrophages.
