Isolation And Purification Of Taiwanofungus Camphoratus Polysaccharides And Its Effect On Macrophage Function

Polysaccharide is a kind of natural Polymeric compound which is composed of aldose or ketose by glycosidic bond.Fungal polysaccharide is a kind of metabolites produced by fungal fruiting bodies and mycelium,which has a wide range of pharmacological activities.The Taiwanofungus camphoratus is a precious medicinal fungus unique to Taiwan,which has anti-tumor,hepatoprotective and antiinflammatory effects.As an important active component in antrodia,the effect of polysaccharide on macrophage function remains unclear.The biological activity of polysaccharide is closely related to its molecular weight,chemical composition,and the structure characteristics of sugar residues.The sources,extraction and purification methods of polysaccharides are directly related to the kinds of polysaccharides obtained.In this study,the optimal extraction conditions of antrodia polysaccharide were optimized,and a kind of antrodia polysaccharide with uniform molecular weight was isolated and purified to investigate its effect on the function of macrophages.In this paper,the liquid deep fermentation technology was used to expand the culture of antrodia strains and obtain mycelium.After lyophilized mycelium was ground,lipid removal was carried out,and the hot water extraction temperature,time and liquid-solid ratio which affected the extraction rate of polysaccharide were optimized by single factor method.The final result was that the extraction rate of polysaccharide was the highest when the temperature was 90℃,the time was 4h and the liquid-solid ratio was 35:1.Based on the results of single factor optimization,response surface analysis was used to investigate the interaction effects of extraction temperature,time and liquid-solid ratio on polysaccharide extraction,and to determine the best parameters of the three factors.The optimal extraction conditions were obtained: temperature: 93.6℃,time: 4.5h,liquid-solid ratio: 40:1.The optimized conditions were verified,and the results showed that the extraction rate of polysaccharide was 19% higher than that before optimization.After protein removal and alcohol precipitation treatment,the crude polysaccharides of antrodia antrodia were obtained.The crude polysaccharide was separated and purified by cellulose DEAE-52,glucan gel G-25,glucan gel G-50,etc.The purified polysaccharide with uniform molecular weight was screened and named as ACPSA,and the molecular weight and monosaccharide components of ACPSA were determined.The Mp,Mw and Mn of ACPSA were 13.924 KDa,16.088 KDa and11.696 KDa.ACPSA is composed of three sugars,namely galactose,glucose and mannose,with a molar ratio of 0.122:0.835:0.042.Taking ACPSA as the research object,ACPSA was applied to human monocytes(THP-1 cells)and mouse primary bone marrow macrophages(BMDM cells)to explore its effect on monocyte differentiation and macrophage function.The results showed that polysaccharide ACPSA can promote the differentiation of THP-1 monocytes into macrophages,induce M1-type polarization of THP-derived macrophages and mouse bone marrow macrophages,enhance the secretion of IL-6,IL-1β,TNF-α,CD86,i NOS and other pro-inflammatory factors,and inhibit the apoptosis of macrophages.In this paper,the optimum extraction conditions were obtained.A kind of polysaccharide ACPSA with uniform molecular weight was obtained by isolation and purification.The polysaccharide can regulate the function of macrophages and enhance the immune effect.This study can provide some theoretical basis for the development of related products and pharmacological mechanisms.