Extraction Of Hericium Erinaceus Fruiting Body Polysaccharide And Its In Vitro Anti-gastric Cancer Activity

Hericium erinaceus,a well-known edible and medicinal mushroom.It had been demonstrated with a wide range of beneficial activities.The polysaccharides fraction was extracted and purified from the fruiting bodies of H.erinaceus.The characteristics of the polysaccharide glycosidic bonds,functional groups and monosaccharides composition were analyzed.The article choose human gastric cancer cells（SGC-7901）as the research object to explore the anti-tumor activity of H.erinaceus water-soluble polysaccharides and the effect of plus chemotherapy drugs cisplatin（DDP）from the cell proliferation,cell apotheosis,migration.Firstly,the experiment extracted H.erinaceus polysaccharides and the optimal conditions for extraction by orthogonal array were as follows:solid/water ratio was 1:60（W/V）,ethanol concentration of 70%,the extracting temperature was 90℃,the extracting time was 2 h.Under this condition,the repeated experiment derived polysaccharides average extraction rate of 8.37%±0.58%.Whereafter,the protein of the polysaccharides was removed with Sevage method and the protein content was determined by codemus brilliant blue method.The deproteinization rate reached 90.26%±1.94%.The sugar content of H.erinaceus polysaccharides after depolarization was 73.92%±1.26%by phenol sulfuric acid method.H.erinaceus fruiting bodies mixture polysaccharide（HFPM）was further purified by DEAE-52 ion exchange chromatography,and four ablution peaks were obtained.HFPM2 had the most significant inhibitory effect on SGC-7901,and the IC₅₀value of 48 h was 626.52μg/mL±2.32μg/mL.Therefore,HFPM2 was purified in the next step.H.erinaceus fruiting bodies polysaccharide（HFP）were available for the performed to obtain homogeneously purified polysaccharides for the Sephadex G-200gel filtration column chromatography.Next,the components of HFP were investigated.No protein content was detected in HFP.The total sugar content of HFP was 91.05%±0.16%.The glucuronic acid content was 4.26%±0.23%,which was an acid polysaccharides.In this article,HFP were hydrolyzed by trifluoroacetic acid method and precolumn derivation of PMP.The monosaccharides were analyzed by capillary electrophoresis.HFP was composed of four monosaccharides,namely Xyl,Rha,Glc and Glu A at a molar ratio of 2.8:2.2:5.1:1.0.Finally,the effect of HFP on SGC-7901 cells was investigated.HFP could effectively inhibit the proliferation of SGC-7901 cells.The inhibitory effect of HFP on SGC-7901 was time-dose dependent,with significant difference at 50μg/mL and extremely significant difference at 200μg/mL.At 48 h,the IC₅₀0 value was 361.78μg/mL±5.49μg/mL.It also had significant effects in combination with low dose DDP.Through observation of morphology,detection of Caspase activity,and exploration of Bax and Bcl-2 expression.It was found that HFP could cause apoptosis of SGC-7901cells.The experiment results show that the control cells within 24 h was easier to achieve healing,and the experimental group cells was dose dependent inhibition of scratches closed.HFP can effectively inhibit the SGC-7901 cells mRNA expression of MMP-2,9.Therefore,HFP can effectively inhibit cell migration.In this paper,the H.erinaceus fruit-body polysaccharides was systematically studied,which proved that the H.erinaceus polysaccharide had good anti-gastric cancer activity,and was a promising development prospect.