Empirical Study Of Anti-tumor Effect Of Carboxymethylation Schizophyllan Polysaccharide

Background and Objective:With the incidence of cancer rising,as people today are having increased chances of getting a malignant tumor,the crisis is now the most important medical task. currently the most popular ways of treating malignant tumor are:operation,radiation and medicinal,medicinal these three ways are the most popular but they are not the best.so scientists today are trying to find the "best" way to treat tumors.Normal immune system can recognize and kill cancer cells,immune protection is poor or not normal cells to identify the poor,can lead to increased incidence of cancer.Therefore,the immune stimulation of immune regulator and is considered a potential Kangai Ji.Polysaccharide is an important biological response modifier.Polysaccharide as the promotion of an immune T cells can not only promote,B cells,NK cells and macrophages of the immune function.Can promote the synthesis of cytokines,or immune cells to increase the sensitivity of cytokines.Can be used to treat cancer, there is no bone marrow suppression,and other toxic.Plant polysaccharides by the tumor Immunoregulation of the body to the anti-tumor effects have been recognized by the world.In order to enhance the immune regulation of polysaccharide,the polysaccharide may have to modify some of the new physical and chemical properties,is conducive to the QSAR on,to increase its anti-tumor effect,is one of polysaccharide on a hot,now Application of a variety of methods have been modified and made great progress.Schizophyllan of Exopolysaccharide(Schizophyllan,SPG) is a medicinal fungus from Schizophyllan bacteria(Schizophyllum communeFries) deep fermentation produced a neutral exopolysaccharide,aβ-(1-6) glucosidase branchβ-(1-3)-D glucan structure,the main chain every three units of glucose produced a branch.In addition it has a good anti-tumor activity,for the food industry,chemical industry,such as the characteristics.Schizophyllan polysaccharide(SPG) immune function as a regulator,mainly through enhancing the body's immune function to kill or inhibit tumor cells.Their role at home and abroad immune mechanism of a number of reports.Such as in vitro,10-100μg/mL dose of SPG can improve concanavalin A-induced mice splenocyte proliferation.Antagonism immunosuppressant hydrocortisone on the lymphocyte proliferation and increase anti-sheep spleen lymphocytes the number of red blood cell antibody,the skin and can increase the delayed-type hypersensitivity reactions,improve cellular immune function.The experimental mice,Schizophyllan polysaccharide on the S-180 of the ascites and solid tumors,Lewis lung cancer,and other anti-tumor activity demonstrated.Polysaccharide immune activity by various factors,including the solubility, molecular weight,chemical structure,route of administration and dose,extraction methods and animal species,such as by the drug.Schizophyllan polysaccharide has been proved to be a good anti-tumor and anti-infection,enhance immunity,such as role.However,domestic production Schizophyllan products there is a soluble polysaccharide poor,large molecular weight,the effect not sure,and so on. Carboxymethylation as a common chemical reaction in starch,cellulose,such as the modification of polymer materials are widely used.In recent years,studies have shown that a certain degree of substitution of the carboxymethylation polysaccharides are often able to play enhanced role in the anti-tumor activity.This is the first of the larger molecular weight polysaccharides Schizophyllan carboxymethylation structure of the different degree of substitution of the modification,to a smaller molecular weight,water-soluble,and low viscosity,easy to make various formulations,to facilitate delivery,And the mechanism of a preliminary study,raising hope that through the modification of its anti-tumor effect, for the development of new anticancer drugs provide a theoretical basis.Method:1.Preparation carboxymethylation Schizophyllan polysaccharide,was measured using the gel filtration its molecular weight.2.With BLCB/C mice by intraperitoneal cultivation of the establishment of liver cancer model,the establishment of negative control group(NS),the positive control group(5-Fu),Schizophyllan polysaccharide Group(SPG),two different degree of substitution of carboxymethyl Schizophyllan polysaccharide group (CM-SPG1),(CM-SPG2).After administration to the tumor size,rate of tumor, pathological examination,and other indicators before and after the evaluation of modified polysaccharides Schizophyllan role in tumor-bearing mice results.3.To rat liver cancer cells H22 and human lung cancer cell lines GLC-82 for the target,set up to the negative control group,the positive control group, Schizophyllan polysaccharide group and the different degree of substitution carboxymethyl Schizophyllan polysaccharide group,based on trypan blue Resist staining and 4-methyl-唑salt micro-reaction assay(MTT) operation,to cell mortality and OD value before and after the evaluation of modified polysaccharides Schizophyllan role in the tumor cells in vitro results.4.Liver cancer tumor-bearing mice were divided into four groups,a control group (NS),the positive control group(CY) Schizophyllan polysaccharide Group(SPG) and carboxymethylation Schizophyllan polysaccharide treatment group (CM-SPG1 Group).Executed before picking eyeball blood separation medium, measured by RIA serum TNF-α,IL-2 content.According to a statement operation5.SPSS11.0 statistical analysis using statistical software package with actual data to mean±standard deviation((?)±s),(1) at different times,different drug, different doses of OD value compared using 4×3factorial,Poor analysis (Factorial analysis),p＜0.05 pm that there was a significant difference.(2) different drugs,different time points compared groups using single-factor analysis of variance(One-Way ANOVA).February 2 comparison group using LSD(or SNK).p＜0.05 pm that there was a significant difference.(3) different times,different drugs in the mortality rate of multiple lines between the X2 test (Chi-Square Test),p＜0.05 pm that there was a significant difference.(4) different groups of serum determination of a single-factor analysis of variance (One-Way ANOVA).February 2 comparison group using LSD(or SNK).p＜0.05 pm that there was a significant difference.Results:1.Carboxymethyl Schizophyllan molecular weight polysaccharides to 4.5×105 Da2.Tumor-bearing mice results(1) mouse tumor weight of each group-a significant difference(p＜0.01);SPG group and the CM-SPG1 and CM-SPG2 showed significant differences(p＜0.05),modified and modified before the Replace two of polysaccharide Schizophyllan there was a significant difference,modified tumor weight is less than before the modified tumor weight;CM-SPG1 and CM-SPG2 was no significant difference(p＞0.05),two different degree of substitution Carboxymethyl polysaccharide Schizophyllan the inhibitory effect no significant difference.(2) inhibition of Congdadaoxiao each group were:5-Fu,CM-SPG1,CM-SPG2, SPG.(3) organizations pathology of the tumors form similar groups,are poorly differentiated cancer and invasive skeletal muscle,are visible coagulation necrosis. SPG Group and the CM-SPG-1 group of tumor cells relatively dense,5-Fu relatively loose group of tumor cells,were shallow;coagulation necrosis area ratio of 5-Fu largest group,SPG Group followed,CM-SPG-1 smallest group;Number of apoptosis 5-Fu largest group,CM-SPG-1 group followed,SPG Group least-cell vacuoles 5-Fu Group and the CM-SPG-1 group is more,SPG at least,such as inflammatory lymphocytes Infiltration of connective tissue cells found in SPG Group and the CM-SPG-1 group,5-Fu group was not obvious.3.CM-SPG role in liver cancer cells in vitro H22 strain of trypan blue resist staining measured by the test cell mortality X2 each group there was significant difference(p＜0.05).SPG,CM-SPG,NS Group mortality rate was not significantly different(p＞0.05).Before and after the modification that Schizophyllan polysaccharides in vitro at the end of liver cancer cells have shown inhibition.4.CM-SPG role in lung cancer cells in vitro GLC-82 of the MTT OD measured by the value of factorial analysis,that the treatment group and 5-Fu control group there were significant differences(p＜0.05).Compared with normal saline to the treatment group had no significant difference(p＞0.05),before and after modification modified Schizophyllan polysaccharides in vitro no effect on lung cancer cells.5.Serum TNF-α,IL-2 in each group in a significant difference(p＜0.05);SPG, CM-SPG group was significantly higher than the NS Group and CY Group,CY group was significantly lower than the NS group.CM-SPG SPG Group and the Group of significant differences(P＜0.05),CM-SPG1 content higher than the SPG Group.Modified,Schizophyllan polysaccharide enhance immunity effect.Conclusion:1.Schizophyllan carboxymethyl polysaccharides can be modified and reduced its molecular weight.2.Schizophyllan polysaccharide and carboxymethyl Schizophyllan polysaccharides in vitro had no direct role in anti-tumor cells.3.Carboxymethyl Schizophyllan polysaccharides in liver cancer in mice showed anti-tumor effect,the anti-tumor effect than carboxymethylation ago has improved.4.Carboxymethyl Schizophyllan polysaccharides to improve liver cancer in mice TNF-α,IL-2 content,can increase immunity inhibit tumor growth.