Chemical composition of Cyclocarya paliurus polysaccharide and inflammatory effects in lipopolysaccharide-stimulated RAW264.7 macrophage

Abstract

This study was designed to study the chemical composition of Cyclocarya paliurus polysaccharide and inflammatory effects in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage. A new elution (0.3% NaCl aqueous solution) of Cyclocarya paliurus polysaccharide (CPP-3) was characterized by different methods such as fourier transform infrared spectra (FT-IR), UV-vis, gas chromatography-mass spectrometry (GC-MS), high performance gel chromatography (HPGLC) and scanning electron microscopy (SEM). Cell viability was measured by MTT test, phagocytosis assay was measured by Neutral red uptake assay, nitrite was measured by Griess assay, TNF-α and IL-1β analysis were measured by ELISA, PGE₂ was measured by enzyme immunoassay system. The results showed that CPP-3 was comprised of two polysaccharides with average molecular weight (Mw) of 5.69×10⁴Da and 4.94×10³Da. CPP-3 contains six monosaccharides, of which are rhamnose (Rha), arabinose (Ara), xylose (Xyl), mannose (Man), glucose (Glu), galactose (Gal), the molar ratio of six monosaccharides is 0.060:0.109:0.053:0.128:0.293:0.357. CPP-3 increased the amount of NO released from mouse macrophage RAW264.7 and significantly increased the levels of TNF-α, IL-1β and PGE₂ (P<0.01). CPP-3 suppressed LPS-stimulated RAW264.7 macrophage to release NO, TNF-α, IL-1β and PGE₂ (P<0.01). CPP-3 and LPS accounted for synergistic effect on the release of NO and TNF-α, CPP-3 and LPS accounted for antagonistic effect on the release of IL-1β and PGE₂.

Keywords: Chemical composition; Cyclocarya paliurus polysaccharide; Inflammatory effects; LPS-stimulated; RAW264.7 macrophage.