Effect Of Polysaccharide From Arenaria Kansuensis On Immune Function In Sarcoma180 Tumor-Bearing Mice

Objective:The purification of polysaccharides from Arenaria kansuensis was studied by molecular weight determination,monosaccharide analysis and immunological activity.To investigate the immune activation effect of polysaccharide from Arenaria kansuensis on S180 tumor-bearing mice,and to provide basis for further isolate polysaccharides from Arenaria kansuensis,which contains antitumor active components.Methods:1)The crude polysaccharide from Arenaria kansuensis(AKCP)was extracted by water extraction-alcohol precipitation method,and was separated and purified by DEAE cellulose 52 column and Sephadex G-75 column.The total sugar content of polysaccharides was determined by phenol-sulfuric acid method.GPC-MALS and HPLC were used to detect the molecular weight and monosaccharide composition.MTT assay was used to detect the effects of polysaccharides at different stages on the proliferation of spleen lymphocytes in vitro.The levels of supernatant in the culture of abdominal macrophages in mice of AKP-2a concentration groups of IFN-?level were detected by NO release assay and ELISA.2)Kunming mice were randomly divided into 6 groups:normal group,model group,lentinan group,AKCP high,medium and low groups.The model of S180tumor-bearing mice was established,in addition to the normal group.Normal group and model group were given intragastric administration of distilled water 0.5 mL/d.Ientinan group was given 0.25 mg/kg lentinan intraperitoneal injection of 0.1 mL/d.AKCP high,medium and low groups mice were given lavage 200,100,50 mg/kg AKCP respectively.After continuous administration for 8 d,blood was taken from the eyeball and tumor tissue was stripped under aseptic conditions.The tumor tissue inhibition rate,weight gain rate,thymus index,spleen index,splenic lymphocyte proliferation activity,NK cell killing activity and CD3+/CD4+/CD8+subsets were measured.The mRNA expression levels of IL-2,TNF-?,IFN-?in splenic lymphocytes and its serum levels were measured.Results:1)AKCP was separated and purified to obtain AKP-2a with high total sugar content,and its molecular weight range and 10 main monosaccharides were preliminarily determined.Compared with control group,the AKCP,AKP-2 and AKP-2a treatment groups with high,medium and low concentrations and lentinan positive control group all had a promoting effect on the proliferation of spleen lymphocytes by MTT in vitro(P<0.05).By NO release experiment and IFN-?ELISA detection,AKP-2a showed a concentration dependent activation effect on mice peritoneal macrophages in vitro(P<0.01).2)AKCP-H group and lentinan group has obvious tumor suppression effect(P<0.05).Splenic lymphocyte proliferation and NK cells killing activity of AKCP-H group and lentinan group were higher than model group(P<0.05).Compared with the model group,AKCP-H,M,L groups of CD3~+CD8~+significantly increased(P<0.05).The expression of IFN-?mRNA in spleen lymphocytes of AKCP-H group and lentinan group was higher than normal group(P<0.05).Serum levels of IFN-?and IL-2 in AKCP-L group and lentinan group were higher than model group(P<0.05).Compared with the normal group,the serum TNF-?in AKCP-H,M,L groups was significantly increased(P<0.05).Conclution:1)This study obtained AKP-2a polysaccharide through isolation and purification of Arenaria kansuensis,which has a high total sugar content.The multiple monosaccharides and the molecular weight of AKP-2a were identified,and the preliminary results showed that AKP-2a had the biological activities on lymphocyte proliferation andmacrophage function.2)Polysaccharide of Arenaria kansuensis had abilities to activate the immune function of S180 tumor-bearing mice.