Study On Extraction And Antitumor Activities Of Glycosides From Acer Saccharum

Objective This paper aimed the anti-tumor activities of active constituent andmonomeric compound of the Acerraceae, Acer Saccharum were extracted and separatedfor screening the anti-tumor activities.Methods (1) Extraction of glycosides: Acer Saccharum was soaked in75%ethanolsolution for12hour. Extractive were isolated by D101Ultrasonication Extraction andeluted by Macroporous Adsorption Resin. The glycosides were obtained by concentratingthe50%ethanol eluent.(2) Separation of glycosides: The monomeric compound wasisolated by routine methods such as silica gel column chromatography, preparation thinlayer chromatography separation and recrystallization.(3) Structural characterization:The compounds were confirmed by13C-NMR,1H-NMR and MS analysis methods.(4)The anti-tumor activities of active constituent of new ten compounds which isolated fromglycosides were tested by the paper disc method. The cancer cell lines are SG7901gastriccancer and SMMC7721hepatocellular carcinoma cell lines.(5) The anti-tumor activitiesof monomeric compound were confirmed by the paper disc method.(6) To investigate theanti-tumor mechanism of compounds, MTT experiment was utilized for monitoring theinhibition of cell proliferation. In addition, Inverted Microscope applied for the supervisionof apoptosis and Flow Cytometer for monitoring SGC7901cell cycle.Results We systematically isolation seven compounds and three of them exhibitantitumor activity to SMMC7721and SGC7901. The results show that compounds exhibithigh anti-tumor activity to negative control group (P<0.05) when the concentration ofTQ-B3, TQ-G1and TQ-I1were12.5~100μg·mL-1,25.0~100μg·mL-1and50.0~100μg·mL-1,respectively. Inverted Microscope image reveals that the tumor cells have no connectionbetween each other and tumor cell body shrinked in the present ofTQ-B3. Anti-tumor effect of TQ-B3in vivo demonstrated that middle dose and high dosegroup decreased the tumor cell in SMMC7721mice tumor-burdened model to1.237±0.480g and0.958±0.593g, respectively. The middle dose and high dose of TQ-B3 increased the spleen weight of SMMC7721mice tumor-burdened model to0.163±0.043and0.180±0.064g respectively, and the thymus weight to0.199±0.078g,0.214±0.511grespectively. The middle dose and high dose of TQ-B3decreased the tumor weight ofSGC7901mice tumor-burdened model to1.307±0.631g and0.958±0.122g respectively.The middle dose and high dose of TQ-B3increased of SGC7901mice tumor-burdened model to0.161±0.02and0.182±0.064g respectively, and thethymus weight to0.209±0.037g,0.214±0.511g respectively, in which all of the resultshave significant difference by comparison with saline (P<0.05).The result of TQ-B3effects on tumor cell cycle experiments indicate that G0/G1,G2/M and S-phase at2μg·mL-1were49.56±1.4,8.32±0.69and42.12±1.36respectively,and at4μg·mL-1were49.67±1.72,5.55±0.64and44.78±0.94respectively, and at8μg·mL-1were71.67±1.53,0and28.33±0.83respectively.Conclusions The results indicated that compounds TQ-B3, TQ-G1and TQ-I1exhibit anti-tumor activities. The anti-tumor mechanism of TQ-B3displayhighest inhibiting effect to SMMC7721and SGC7901and Dose-Effect Relationship.Compounds can induce tumor apoptosis and S-phase cycle arrest, eventually enhance theimmune capacity.