This study identified two homogeneous acidic polysaccharides from Gardeniae fructus, GJP50-3 and GJP50-4, which exhibited potential immunomodulatory activities in macrophage activation assays, via liquid-chip technology, and in a zebrafish model. Monosaccharide composition analysis and gel permeation chromatography revealed that GJP50-3 and GJP50-4 were composed of Rha, GalA, Glc, Gal, and Ara in specific ratios and had molecular weights of 91.5 kDa and 140.3 kDa, respectively. Based on FT-IR, GC-MS, and NMR analyses, these polysaccharides were identified as typical pectin polysaccharides with methylation degrees of 24.7 % and 21.4 %, respectively. The primary structures of GJP50-3 and GJP50-4 included linear HG domains and branched RG-I domains with arabinans and AG side chains. In vitro, GJP50-3 and GJP50-4 could stimulate NO release and increase the secretion of TNF-α in a RAW 264.7 macrophage model. Luminex liquid suspension chip detection revealed that GJP50-3 significantly promoted the secretion of multiple interleukins [IL-6, IL-9, IL-10, IL-12 (p40), IL-12 (p70), IL-13], TNF-α, and chemokines (G-CSF, GM-CSF, MCP-1 and RANTES). In vivo, these polysaccharides could also increase NO release and neutrophil count in a zebrafish model. These findings suggested that GJP50-3 and GJP50-4 might have the potential to be used as immunomodulators in the food and pharmaceutical industries.
Keywords: Immunomodulatory activities; Pectin-polysaccharide; Structure characteristics.
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