Molecular mechanisms of macrophage immunomodulation mediated by Areca inflorescence polysaccharides based on RNA-seq analysis

The elucidation of the immunomodulatory molecular mechanisms of polysaccharides has contributed to their further development and application. In this study, the effect of Areca inflorescence polysaccharide (AFP2a) on macrophage activation was confirmed and the detailed mechanisms were investigated based on a comprehensive transcriptional study and specific inhibitors. The results showed that AFP2a induced macrophage activation (M1 polarization), promoting macrophage proliferation, reactive oxygen species production, nitric oxide and cytokine release, and costimulatory molecule expression. RNA-seq analysis identified 5919 differentially expressed genes (DEGs). For DEGs, GO, KEGG, and Reactome enrichment analyses and PPI networks were conducted, elucidating that AFP2a activated macrophages mainly by triggering the Toll-like receptor cascade and corresponding adapter proteins (TIRAP and TRIF), thereby resulting in downstream NF-κB, TNF, and JAK-STAT signaling pathway expression. The inhibition assay revealed that TLR4 and TLR2 were essential for the recognition of AFP2a. This work provides an in-depth understanding of the immunoregulatory mechanism of AFP2a while offering a molecular basis for AFP2a to serve as a potential natural immunomodulator. [Display omitted] •Areca (Areca catechu L.) flower polysaccharide (AFP2a) with low concentrations promoted macrophage activation and make it to M1 polarization.•RNA-seq analysis revealed AFP2a on the macrophage mainly activates toll-like receptors and its corresponding downstream signaling pathways.•TLR4 and TLR2 were identified as AFP2a binding receptors through the inhibitor experiment.