Characterization of polysaccharides from Tetrastigma hemsleyanum Diels et Gilg Roots and their effects on antioxidant activity and H₂O₂-induced oxidative damage in RAW 264.7 cells

This work presents an investigation on the composition and structure of polysaccharides from the roots of Tetrastigma hemsleyanum (THP) and its associated antioxidant activity. It further explores the protective effect of THP on RAW264.7 cells against cytotoxicity induced by H₂O₂. Ion chromatography (IC) revealed that THP contained glucose, arabinose, mannose, glucuronic acid, galactose and galacturonic acid, in different molar ratios. Furthermore, gel permeation chromatography-refractive index-multiangle laser light scattering (GPC-RI-MALS) was employed to deduce the relative molecular mass (M_w) of the polysaccharide, which was 177.1 ± 1.8 kDa. Fourier transform infrared spectroscopy (FT-IR) and Congo red binding assay highlighted that the THP had a steady α-triple helix conformation. Similarly, assays of antioxidant activity disclosed that THP had reasonable concentration-dependent hydroxyl radical and superoxide radical scavenging activities, peroxidation inhibition ability and ferrous ion chelating potency, in addition to a significant 1,1-diphenyl-2-picrylhydrazyl radical scavenging capacity. Moreover, THP could protect RAW264.7 cells against H₂O₂-induced cytotoxicity by decreasing intracellular ROS levels, reducing catalase (CAT) and superoxide dismutase (SOD) activities, increasing lactate dehydrogenase (LDH) activity and increment in malondialdehyde (MDA) level. Data retrieved from the in vitro models explicitly established the antioxidant capability of polysaccharides from T. hemsleyanum root extracts.