RESEARCH PAPER

Fermented Flammulina velutipes Polysaccharide Inhibits Macrophage Inflammatory Response through Nuclear Factor-κB-NOD-Like Receptor Family Pyrin Domain-Containing Protein 3 Signaling Pathway

  • MA Sheng ,
  • GAO Qingying ,
  • XU Jianxiong
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  • Shanghai Key Laboratory for Veterinary and Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China

Received date: 2021-07-11

  Online published: 2022-02-15

Abstract

To investigate the effects and mechanism of Flammulina velutipes polysaccharide (FVP) and fermented Flammulina velutipes polysaccharide (FFVP) on the inflammatory response of mouse mononuclear macrophages (RAW264.7), we constructed an inflammatory model of RAW264.7 with lipopolysaccharide (LPS). CON group (normal medium), LPS group (normal medium+1 μg/mL LPS), FVP group (normal medium+1 μg/mL LPS+25, 50 or 100 μg/mL FVP) and FFVP group (normal medium+1 μg/mL LPS+25, 50 or 100 μg/mL FFVP) were set, cell viability, phagocytosis ability, the contents of reactive oxygen species (ROS) and nitric oxide (NO) as well as the contents and mRNA relative expression levels of inflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-18 (IL-18) and tumor necrosis factor-α (TNF-α) in RAW264.7 were determined to compare the inhibitory effects of FVP and FFVP on the inflammatory response of macrophages. RAW264.7 were treated with nuclear factor-κB (NF-κB) inhibitor BAY11-7082, and the protein relative expression levels of phosphorylated nuclear factor-κB inhibitor α (p-IκBα), NOD-like receptor family pyrin domain-containing protein 3 (NLRP3), cysteinyl aspartate specific proteinase-1 (Caspase-1) and IL-1β were detected by Western blot to explore the mechanism of FVP and FFVP on LPS-induced inflammatory response of macrophages. The results showed that both FVP and FFVP could inhibit the increase of ROS and NO contents in RAW264.7 induced by LPS, and the difference was significant at 100 μg/mL (P<0.05). Compared with LPS group, FVP and FFVP increased phagocytosis ability of RAW264.7 by 37.65% and 47.06%, respectively. FVP and FFVP decreased the contents and mRNA relative expression levels of inflammatory cytokines IL-1β, IL-6, IL-18 and TNF-α in a dose-dependent manner. Western blot results showed that the protein relative expression levels of p-IκBα, NLRP3, Caspase-1 and IL-1β in RAW264.7 were significantly decreased by BAY11-7082 combined with FVP or FFVP (P<0.05), indicating that FVP/FFVP could inhibit the activation of NLRP3 signaling pathway by reducing IκBα phosphorylation. In conclusion, both FVP and FFVP can enhance cell viability and phagocytosis ability of RAW264.7, reduce ROS and NO contents, and inhibit macrophage inflammatory response by inhibiting NF-κB-NLRP3 signaling pathway activation. The anti-inflammatory effect of FFVP is better than that of FVP at the same concentration.

Cite this article

MA Sheng , GAO Qingying , XU Jianxiong . Fermented Flammulina velutipes Polysaccharide Inhibits Macrophage Inflammatory Response through Nuclear Factor-κB-NOD-Like Receptor Family Pyrin Domain-Containing Protein 3 Signaling Pathway[J]. Chinese Journal of Animal Nutrition, 2022 , 34(2) : 1205 -1216 . DOI: 10.3969/j.issn.1006-267x.2022.02.052

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