Isolation, Structural Analysis and Anti-Inflammatory Activity of a Polysaccharide from Ilex cornuta Fruits.

Abstract

In the present study, a polysaccharide from Ilex cornuta fruits (LCFP-3) was obtained by hot water extraction, Diethyaminoethyl cellulose-52 (DEAE-52) chromatography column and Sephadex G-100 gel column purification. Its structural characteristics were further explored using high performance anion exchange chromatography (HPAEC), gas chromatography and mass spectrometry (GC/MS), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. Monosaccharide composition analysis revealed LCFP-3 contained mainly Galactose (31.92 %), Arabinose (25.87 %) and Galacturonic acid (23.35 %) while small percentage of Rhamnose, Glucose, Mannose and Xylose. Chemical composition analysis showed that the total sugar content of LCFP-3 was 90.31 % and the protein content was 0.246 %. Gel permeation chromatography (GPC) analysis showed that its average molecular weight was 41.199 kDa. Structural analysis showed that LCFP-3 may be composed of residues, T-α-Arap, T-α-Rhap, 1,3-α-Arap, 1,4-α-Arap, T-β-Galp, 1,4-α-GalpA(OMe), 1,4-β-Glcp, 1,3-β-Galp, 1,3,6-β-Manp, 1,6-β-Galp, 1,3,4-β-GalpA, 1,4,6-β-Manp, 1,3,6-β-Glcp, 1,2,3,4-α-Xylp. The anti-inflammatory activity of LCFP-3 was evaluated using lipopolysaccharide (LPS)-induced RAW246.7 macrophages. The results showed that 1-200 μg/mL LCFP-3 could dose-dependently protect against LPS-induced toxicity and 1 μg/mL LCFP-3 could significantly inhibit LPS-induced NO production. Therefore, LCFP-3 exerted an anti-inflammatory activity and has great potential as a functional ingredient.