Study On Extraction,Isolation And Pharmacological Activity Of Polysaccharides From Trametes Versicolor(L.) Lloyd

Trametes versicolor?L.?Lloyd originating from the virgin forest of China is a fruiting body of Trametes versicolor?L.?Lloyd.Polysaccharide is the main effective part of Trametes versicolor?L.?Lloyd,which has biological activities.The medicinal products of the polysaccharides of Trametes versicolor?L.?Lloyd are rare on the market,and the most commonly used product is Yunzhi Gantai,which is used as an immunomodulator to treat acute and chronic active hepatitis.However,there are few studies on its anti-hepatocarcinoma activity and anti-drug liver injury.Based on the premise of this study,the activities of the polysaccharide of Trametes versicolor?L.?Lloyd were studied and their mechanisms were explored.In this context,the polysaccharides were extracted by water,pH=2 hydrochloric acid solution,6%urea solution,3%trichloroacetic acid solution and 0.2M sodium hydroxide solution.The polysaccharides obtained were respectively named as YZ₀,YZ₁,YZ₂,YZ₃ and YZ₄.Then Sevage method was used to remove the proteins from the polysaccharides,and the activated carbon method was used to decolorize the polysaccharides.Finally,the purified polysaccharides were obtained.The contents of neutral sugar,protein and glucuronic acid in YZ₁ were determined by colorimetry method.The composition of polysaccharide monosaccharide was determined by gas chromatography with acrylonitrile acetate derivatization.HPSEC was used to determine the composition of YZ₀.And the antitumor activity of various polysaccharides and the protective effect of YZ₀ polysaccharides on liver injury induced by APAP were studied.Groups were set up to investigate the antitumor effect of acid-soluble polysaccharides from Trametes versicolor?L.?Lloyd with the model of H₂₂2 hepatoma transplanted tumor mice,once a day and continuous administration for 10 days,including normal group,model group,CTX group,and the high and low dose group of polysaccharides from Trametes versicolor?L.?Lloyd.Measure the indexes included inhibitory rate,organ index of liver,thymus and spleen and the levels of serum included IL-2,IL-6,IFN-?,TNF-?,AST,ALT,BUN and CRE.Finally,the morphological changes and apoptosis of tumor tissues after H&E staining were observed under light microscope,and the expression of Bcl-2,Bax,Caspase-3 and Caspase-9 were detected by immunohistochemistry and Western blot in mice with high and low dose of YZ₀ to further study the anti-tumor mechanism of polysaccharides.The results showed that the polysaccharides could inhibit the growth of tumor tissue to a certain extent,and the high dose group of YZ₀ polysaccharide had the best effect,and the inhibitory rate was 68.3%,which was higher than that of CTX group.And improved the immune organ index and the level of serum cytokine IL-6,IL-2,IFN-?,TNF-?and reduce serum ALT,AST,BUN and CRE levels.The results of H&E staining and apoptosis detection in tumor tissue showed that the polysaccharide could promote the apoptosis of tumor cells.Western blot and immunohistochemical analysis showed that the polysaccharides could regulate the expression of related apoptotic proteins and then play an anti-tumor role.The model group was established by APAP-induced liver injury in mice,and the control group,model group and the polysaccharide-treated YZ₀ group were divided into high and low dose groups,administered once daily for 7 days.The protective effect of YZ₀ polysaccharides on liver was evaluated by the determination of ALT,AST,IL-6,TNF-?,SOD,GSH,CAT and MDA and the detection of H&E staining and Western Blot.The results showed that the pretreatment with the polysaccharide of YZ₀ could decrease the increasing level of serum cytokine IL-6,IL-1?,TNF-?,AST and ALT and the content of MDA in liver homogenate,and increase the activity of SOD,CAT and GSH in liver homogenate.H&E results showed that APAP-induced hepatocellular inflammatory infiltration could be ameliorated.Western Blot results showed that pretreatment with YZ₀ could significantly decrease APAP-induced phosphorylation of JNK and ERK phosphorylation.In conclusion,the polysaccharide of Trametes versicolor?L.?Lloyd had good anti-tumor activity and liver protection.The polysaccharide of Trametes versicolor?L.?Lloyd can enhance the body's immune system,regulate the body's inflammatory factor content and the expression of apoptotic proteins,and promote the apoptosis of tumor cells to exert its anti-tumor effect.It also protects the liver from drug toxicity by regulating the antioxidant function,inflammatory factors and phosphorylation of JNK and ERK.