Polysaccharides of Floccularia luteovirens regulate intestinal immune response, and oxidative stress activity through MAPK/Nrf2/Keap1 signaling pathway in immunosuppressive mice

This study explores the novel immunomodulatory effects of polysaccharides from the rare Floccularia luteovirens, a fungus with significant potential yet unexplored bioactive components, traditionally used in Tibetan medicine. This study employs a wide array of analytical techniques, including HPGPC, HPLC, western blotting, ELISA, and 16S rRNA gene sequencing, to comprehensively investigate FLP1's effects. The main structure of FLP1 was characterized by IF-TR and NMR spectrometry. The structural backbone of FLP1 was -*3,6)-(3-D-Glcp-(1 -* and -*2,3)-alpha-D-Manp-(1-*. After immunosuppressed mice treated with FLP1, the findings demonstrated that FLP1 stimulated the production of secretory sIgA and secretion of cytokines (IL-4, TNF-alpha, and IFN-gamma) in the intestine of Cy-treated mice, resulting in the activation of the MAPK pathway. Additionally, FLP1 protected oxidative stress by triggering Nrf2/Keap1 pathways and antioxidation enzymes (SOD, MDA, T-AOC, CAT, and GSH-Px). It also enhanced the intestinal barrier function by regulating the villous height ratio and expression of tight-junction protein. Furthermore, FLP1 remarkably reversed the gut microbiota dysbiosis in immunosuppressed mice by increasing the abundance of Oscilliospiraceae, and Lachnospiraceae, and altered the fecal metabolites by increasing LysoPE (0:0/18:0); 0:0/16:0; 18:1(11Z)/0:0, LysoPG (16:0/0:0), LysoPG 18:1 (2n) PE (14:0/20:1), echinenone, 2-(2-Nitroimidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl) acetamide, and suberic acid which is closely related to the immunity function. These results suggested that FLP1 may regulate the intestinal immune response by modulating the gut microbiota and fecal metabolites in immunosuppressed mice thereby activating the immune system.