Extraction,purification,structural characterization of polysaccharide from Aconiti Lateralis Radix Praeparata and its biological evaluation on immune activity of RAW 264.7 cells
Objective To characterize the chemical structure of polysaccharide from Aconiti Lateralis Radix Praeparata(ACP)and evaluate its immunological activity on RAW 264.7 cells.Methods Crude polysaccharide from Aconiti Lateralis Radix Praeparata was extracted by hot water extraction and the alcohol precipitation method,and purified by the anion exchange chromatography column and the gel chromatography column to obtain homogeneous ACP.The homogeneity and molecular weight distribution of ACP were determined by high-performance gel permeation chromatography(HPGPC),the monosaccharide composition was determined by ion chromatography,and the glycosidic bond type and primary structure of ACP were elucidated by methylation experiments and NMR spectroscopy.Finally,the effects of ACP on macrophages were determined by in vitro cell experiments.Results and conclusions ACP was successfully isolated from Aconiti Lateralis Radix Praeparata with a molecular weight of 11.37 kDa.The ACP was mainly composed of glucose,and the backbone of ACP was →4)-Glcp-(1 → glycosidic bond as the main chain,and the terminal Glcp-(1→ as the brached chain through O-6 linkage of →4,6)-Glcp-(1 → linked to the main chain.The in vitro experiment results showed that ACP promoted the secretion of inflammatory factors including IL-1β,IL-6,iNOS,and TNF-α within a certain concentration range,indicating the immunological activity of ACP.
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