Study On The Protective Effects Of Polysaccharides From Thelephora Ganbajun On Alcoholic Liver Injury In Mice

Thelephora ganbajun is also called Hydrangea bacteria,fungus flower,horse teeth bacteria,belongs to Mycobionta,Hymenomycetes,Polyporales,Thelephoraceae.It is a rare wild medicinal and edible fungus.Studies indicate that Thelephora ganbajun contains polysaccharides,polyphenols,proteins and other bioactive substances with high nutritional value.Therefore,the present study is to investigate the protective effect of Thelephora ganbajun polysaccharide on alcoholic liver injury from the following three aspects.1.The extraction,purification and component verification of Thelephora polysaccharide.This test use ultrasound-assisted water extraction and alcohol precipitation method,the use of single-factor and response surface experiment investigated solid-liquid ratio,extraction temperature,extraction time,ultrasonic power on extraction of Thelephora ganbajun,the optimum extraction conditions of crude polysaccharide from Thelephora ganbajun(CPTG)is achieved.After deproteinization using the method of Sevag,the CPTG become the refined polysaccharide from Thelephora ganbajun(RPTG).Purified polysaccharide from Thelephora ganbajun(PPTG ?)was obtained from RPTG by DEAE-Cellulose52 and Sepharose 4B.Then the composition of PPTG? is analyzed by thin layer chromatography and high performance liquid.The results of the optimum extraction conditions of CPTG showed that solid-liquid ratio was 38:1,extraction time was 3 h,extraction temperature of 89?,ultrasonic power 603 W,the polysaccharide yield was 5.96%;the UV scanning spectrum displayed that PPTG ? was free of nucleic acids and proteins;the IR scanning spectrum suggested that polysaccharide molecules containing pyran ring carbon skeleton and other typical features of polysaccharide groups;thin layer chromatography and high performance liquid verified that Thelephora ganbajun is a heteropolysaccharide made of galactose,xylose and glucose.2.Discussion on the improving effect of Thelephora polysaccharide on alcoholic liver injury from the perspective of the antioxidant,scavenging free radicals and the ability of antioxidation in vivo were studied,while scavenging rate of ·OH?·O2-?·DPPH and reducing power were also determinated.Results:CPTG,RPTG have certain antioxidant effect,the higher the purity and concentration of polysaccharides,the better antioxidant effects they were.Investigate the protective effect of RPTG by establishing acute alcoholic hepalic injury mode.The mice were randomly divided into control group,model group,positive control group(150 mg/kg.bw),RPTG groups(100,200,400 mg/kg.bw).Each group has 11mice,the mice were intragastric administration for 30 days.The model of acute alcoholic liver injury was established on the 31st day,all mice were given 50%alcohol by intragastric administration,except the control group,after 12 h,remove dead mice,10 mice from each group were taken eyeballs and serum was separated.Mice were sacrificed by dislocating,determinate the antioxidant indicators of liver and serum.Results:Compared with the model group,in the RPTG groups,the level of superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px)and valley glutathione(GSH)were significantly increased,content of malondialdehyde(MDA)decreased significantly,and there is a dose-effect concentration of RPTG,wherein the degree of improvement in the high dose group and even some of the indicators have reached the level of the positive control group.Thelephora ganbajun polysaccharide not only has good antioxidant capacity,but also enhance the body's antioxidant enzyme activity.It can inhibited lipid peroxidation.It has a protective effect on alcohol-induced liver injury.3.Taking RPTG as test materials,determined the level of alanine aminotransferase,aspartate aminotransferase(AST),triglyceride(TG)in mouse liver and serum,Observated paraffin section and ultrastructure of mouse liver by the establishment of acute alcoholic liver injury model.the results show:compared with model group,in the RPTG groups,the level of ALT,AST,TG were significantly reduced in liver and blood(P<0.01).In the high-dose group,the vitalities of ALT,AST in serum have reached the positive control group.Pathological section indicated that:In the model group,liver tissue structural disorder,central venous expand and severely deformed,hepatocellular necrosis,inflammatory cell infiltration serious,in the RPTG groups,mice hepatic cord disorders,inflammation and other symptoms of liver cells have different levels of amelioration,the degree of amelioration is more obvious with the more dose;The ultrastructure of liver cell showed that:in the model group,nuclear and mitochondrial deformed severely,nuclear membrane was irregular,mitochondria was blurred,endoplasmic reticulum(ER)was expanded seriously,ribosomal particle shedded,lots of lipid droplets appeared in vision,in RPTG groups,These phenomena have varying degrees of amelioration,the degree of amelioration is more obvious with the more dose.The results suggest that,RPTG can effectively reduce the extent of damage to liver tissue,alcohol-induced acute liver injury in mice showed a protective effect.