| Deerskin,as a traditional Chinese medicine,has been used for a long time.It can treat various diseases such as ulcer,leucorrhea of women,deficiency of kidney and slippery essence.Deerskin glue is made from the skin of sika deer or red deer.It has many functions,such as supplementing qi,blood and kidney.Its efficacy is recorded in compendium of Materia Medica and records of Sichuan Traditional Chinese medicine.Modern research has proved that deerskin gum can replenish blood and enhance immunity.At present,the research on deerskin gum mainly focuses on its functions of blood replenishing,immune regulation and improving serum testosterone,but there is no report on the edible safety of deerskin gum.In addition,the research on the composition of deerskin gum mainly focuses on amino acids,trace elements and other small molecules,while the research on polysaccharide,protein and other major biological macromolecular components is relatively less.Therefore,this study carried out a systematic toxicological evaluation of deerskin gum,on the basis of which a simple and convenient extraction process of deerskin gum polysaccharide was determined,and the purity and structure of the extracted polysaccharide components were analyzed;at the same time,the antioxidant activity of deerskin gum polysaccharide and its protective effect on alcoholic liver injury were studied.The implementation of this study has important practical significance for the comprehensive development and utilization of deerskin gum and its promotion in the field of health food.The contents and results of this experiment are as follows:1.Firstly,the toxicity of deerskin gum was evaluated by acute toxicity test,subchronic toxicity test,micronucleus test of bone marrow cells,sperm deformity test and Ames test.The results of toxicity evaluation of deerskin gum showed that in the acute toxicity test,the oral LD50 of deerskin gum was 10.00 g/kg the subchronic toxicity test,there was no significant difference in blood routine test,blood biochemical index,organ coefficient and histopathology between each dose group and the control group.The micronucleus rate of male and female bone marrow cells did not increase when the dose was less than 10 g/kg.The results of sperm deformity test showed that there was no sperm abnormality in each dose group,and Ames test showed that 5 doses of deer skin glue sample had no mutagenic effect on 4 strains of histidine deficient Salmonella typhimurium.It can be seen that deer skin glue sample was nontoxic and had certain edible safety.2.The extraction process of Polysaccharide from dehskin gum was optimized by water bath extraction combined with orthogonal test.Subsequently,DEAE-52 cellulose column chromatography was used to analyze the polysaccharide and then the molecular structure was analyzed by Sephadex G-200 gel column chromatography and infrared spectroscopy.The optimum extraction conditions were 85?,2.5 hours,25 times of the ratio of material to liquid.The yield of polysaccharide was 19.3%.The polysaccharides extracted by water bath extraction were successively added to DEAE-52 anion exchange column and Sephadex G-200 gel column for further purification,followed by phenol sulfuric acid method.The purified polysaccharide was white floc with purity of 89.4%.Through the reaction of iodine and potassium iodide,it is proved that the polysaccharide does not contain starch,phenols,free amino acids and proteins,and the end has no reducing sugar residue.The polysaccharide contains uronic acid and is acid polysaccharide.Through the analysis of infrared spectrum,it is inferred that the polysaccharide is pyranose.3.To evaluate the antioxidant effect of deerskin gum polysaccharide.In the aspect of antioxidation in vitro,the scavenging effects of·OH radicals,O2-·radicals and the reduction ability of Fe3+were detected.In vivo antioxidant test,D-galactose was injected subcutaneously to establish aging model,and corresponding drugs were given to mice by gavage.The activity of SOD,GSH-Px and the content of MDA in liver and serum were measured after 6 weeks.The results of antioxidant activity test in vitro showed that:there was a certain reducing power and there was a dose-dependent relationship between it and reducing power?the dose was set at 1,2,3,4,5 mg/m L?.Deerskin gum polysaccharide has a strong·OH and O2-·scavenging effect.In vivo antioxidant study of deerskin gum polysaccharide showed that the weight of mice in each experimental group was lower than that in the normal group,and the weight of mice in the model group was the lowest as a whole.Each dose group showed significant difference?P<0.05?and extremely significant difference?P<0.01?in different time periods.There was no significant difference?P>0.05?between the low dose group and the middle dose group in the effect of deerskin gum polysaccharide on the serum of aging mice.The SOD activity of the high dose group increased significantly?P<0.01?,the MDA content decreased significantly?P<0.05?,and the GSH-Px activity had no significant difference?P>0.05?.4.To study the protective effect of deerskin gum polysaccharide on acute alcoholic liver injury in mice.In the last experiment,50%ethanol solution was given to the low,middle,high dose group and the positive control group.Finally,the activity of serum ALT,AST,GSH-Px,SOD and the content of MDA were measured.The mouse liver was made into paraffin and ultra-thin sections,and the rest of the liver was preserved for determination.The indexes of antioxidant damage were MDA content,GSH-Px and SOD activity.The experimental results showed that compared with the model group,the activities of SOD and GSH-Px in the blood of the mice in the different dosage groups and the biphenyl diester positive group increased significantly?P<0.05,P<0.01?,the activities of ALT,AST and MDA decreased significantly?P<0.05,P<0.01?,There was no significant difference?P>0.05?in the low-dose group,MDA content and SOD activity in the middle dose group?P<0.05?,MDA content and SOD activity in the high-dose group were significantly increased?P<0.01?,GSH-Px activity was significantly different?P<0.05?.After acute alcoholic liver injury in mice,the liver cells of each group showed a good improvement effect,and the effect of high dose group was the strongest. |
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