Preparation,Structure Characterization And Antitumor Activity Of Polysaccharide And Selenium Polysaccharide From Coriolus Versicolor

Coriolus versicolor is a kind of traditional Chinese medicine included in“Chinese Pharmacopoeia 2015”.The polysaccharides contained in it have a variety of biological activities,such as reducing blood sugar,anti-tumor and improving immunity.In recent years,it has been found that the synergistic combination of organic selenium and polysaccharides can show higher biological activity.In this paper,polysaccharides from fruiting bodies of Coriolus versicolor were extracted,isolated and purified,and their structure characterization and antitumor activity in vitro were studied.In addition,the antitumor activities of selenium polysaccharides from fruiting bodies of Coriolus versicolor and Coriolus versicolor mycelium were compared and analyzed.The main results are as follows:（1）In this study,crude polysaccharides were obtained from the fruiting body of Coriolus versicolor by degreasing,water extraction,alcohol precipitation,deproteinization and dialysis with a yield of 1.75%.The crude polysaccharides were further purified by cellulose column chromatography and dextran column chromatography to obtain homogeneous polysaccharide YZP-1a.The average molecular weight of YZP-1a was 13884 Da.YZP-1a comprises five monosaccharides:glucose,fucose,galactose,mannose,glucuronic acid,with a mass fraction ratio of1.00:0.133:0.378:0.127:0.030.Through further characterization and analysis,the basic skeleton structure of YZP-1a was as follows:the main chain was composed of→6)-α-D-Glcp-（1→6）-α-D-Glcp-（1→3）-β-D-Galp-（1→6）-β-D-Glcp-（1→[4）-α-D-Galp-（1]₂→4）-α-D-Glcp-（1→.Linked with the O-6 and O-4 position of→4,6）-α-D-Glcp-（1→of the main chain were branches→1）-β-D-Galp-（3→1）-α-D-Glcp and→[1)-α-D-Galp-（4]₃→1）-β-D-Glcp-（6→1）-β-D-Galp-（3→1）-α-D-Galp-（3→1）-β-D-Glcp-（6→1）-β-D-Galp-（3→1）-α-D-Glcp.Linked with the O-6 and O-3 position of→3,6)-α-D-Galp-(1→of the main chain were branchesα-D-Glcp-（1→and→1）-β-D-Glcp-（6→1）-α-D-Glcp.The results of DLS,SEM and TEM showed that the YZP-1a powder was flaky and connected by multiple spheres.A single sphere was easy to aggregate into an irregular chain in aqueous solution,and the average particle size of a single sphere was about 20±5nm.（2）An in vitro cell model was established to explore the anti-tumor activity of YZP-1a.CCK-8 test and scratch test showed that YZP-1a could reduce the cell viability and cell migration ability of A549 cells,HCT116cells and Hep G2 cells in a dose-dependent manner,especially on A549cells.Through AO/EB staining and flow cytometry,it was found that YZP-1a could promote the apoptosis of A549 cells in a dose-dependent manner.At the same time,the results showed that YZP-1a did not show the inhibitory effect on normal cells at the maximum concentration.（3）Selenium polysaccharides Se-YZP-1a in fruiting body of Coriolus versicolor and Se-YZMP-1a in mycelium of Coriolus versicolor were obtained by chemical synthesis and biotransformation.The selenium contents were 173.8μg/g and 30.57μg/g,respectively.Using CCK-8experiment,it was found that Se-YZP-1a with higher selenium content showed stronger cytotoxicity to HCT116 cells.The experiment was carried out on NCM460 cells under the same conditions,and Se-YZP-1a with high selenium content had certain cytotoxicity to NCM460 cells.