Structural characterization of a polysaccharide from Scutellaria baicalensis Georgi and its immune-enhancing properties on RAW264.7 cells.

A novel polysaccharide SPS01-2 (87.5 kDa) was isolated from the roots of Scutellaria baicalensis Georgi. Monosaccharide composition revealed that SPS01-2 consists of rhamnose, arabinose, galactose, galacturonic acid, and glucuronic acid in ratio of 4.4: 67.1: 22.2: 6.3: 1.2. Further investigations using methylation, NMR, and mass spectrometry indicated that SPS01-2 is classified as a type II arabinogalactan (AG-II) with a minor presence of type I rhamnogalacturonan (RG-I). The core structure alternates between 1,2/1,2,4-α-L-Rhap and 1,4-α-D-GalpA, with branches including 1,3,6-β-D-Galp, 1,3-β-D-Galp, T-β-D-Galp, and T-α-L-Rhap. The RG-I regions are linked to 1,6-β-D-Galp, and 1,3,6-β-D-Galp units. Numerous arabinan branches, featuring multiple branching points, are attached to the O-3 position of galactose. Additionally, T-β-D-Galp, 1,6-β-D-Galp, and T-β-D-4-OMe-GlcpA are also linked to galactose in the backbone. Furthermore, SPS01-2 demonstrated potential immune-enhancing properties by dose-dependently increasing proliferation, phagocytosis, and the production of nitric oxide and cytokines (TNF-α, IL-6, and IL-1β) in RAW264.7 cells. It also enhanced the expression of CD80, CD86, and MHC-II at concentrations ranging from 5 to 200 μg/mL. Moreover, the immunostimulatory activity of SPS01-2 was significantly reduced when branch linkages were removed through partial acid hydrolysis. Our findings indicate that SPS01-2 could serve as a natural immunostimulant in the food and pharmaceutical sectors.