RESUMEN
Objective: To extract, separate and purify homogeneous polysaccharides from Rubus sachalinensis and study on monosaccharide component and immunomodulatory activity. Methods: The crude polysaccharides of Rubus sachalinensis (RSP) were extracted by hot water. The Cellulose DE-52 and Sephadex G-100 columns were used to separate and purify homogeneous polysaccharides. The relative molecular mass was analyzed by high-performance gel permeation chromatography, and the monosaccharide composition and structure were preliminarily identified by GC, IR and NMR. The effects on proliferation function of mice spleen lymphocyte proliferation were determined by CCK-8, and the effects on the release capacity of IL-2, IFN-γ and TNF-α were determined by the ELISA kit. Results: Two homogeneous polysaccharides, RSP1-1 and RSP1-2, were separated and purified, with molecular weights of 13 227 and 9 343 determined by HPGPC. They mainly contained arabinose, mannose, glucose and galactose, with the mole ratios at 9.5:7.0:10.3:18.6 and 5.7:11.1:10.3:14.2, respectively. The structure of RSP1-1and RSP1-2 was analyzed by IR and NMR, and RSP1-1 might mainly contain α-1,3-Ara, β-1,4Gal, β-1,6-Glc, β-1,3-Man, and RSP1-2 might mainly contain β-1,4-Gal, β-1,6-Glc, β-1,3-Man. At 5-200 μg/mL, RSP1-1and RSP1-2 stimulated proliferation of spleen lymphocytes (P < 0.05) and promoted lymphocytes to secrete IFN-γ and TNF-α. At 5 μg/mL, RSP1-1and RSP1-2 promoted lymphocytes to secrete IL-2. Conclusion: RSP1-1and RSP1-2 are natural homogeneous polysaccharides that are obtained from this plants for the first time. Its purity and structure were further characterized by IR and NMR. These two homogeneous polysaccharides promoted the proliferation of splenic lymphocyte in different degrees and promoted lymphocytes to secrete IL-2, IFN-γ and TNF-α that all possessing immunomodulating activity.
RESUMEN
Objective: To establish an HPLC fingerprint spectrum of Mongolian medicine Rubus sachalinensis. Methods: An HPLC was performed on the column of phenomsil ODS (250 mm × 4. 6 mm,5 μm) with the mobile phase of 0. 3% phosphoric acid-MeOH with gradient elution at a flow rate of 1. 0 ml·min-1 ,the detection wavelength was 290 nm, the column temperature was 25 ℃and the injection volume was 20 μl. Totally 7 batches of Rubus sachalinensis from different habitats were analyzed. Results:The RSD of relative retention time of the common peaks of Rubus sachalinensis was less than 3%, and that of relative peak areas was below 3%as well, which were in accordance with the requirements of fingerprint. Conclusion:The established HPLC fingerprint has promising accuracy, repeatability and stability, which can be used as one basis for the quality control of Rubus sachalinensis.