The Extraction, Purification, Characterization And Anti-oxidation And Hypoglycemic Activity Of Epimedium Polysaccharide And Glycoprotein

Epimedium brevicornu,a famous Chinese edible herb,was extensively applied to health food and pharmaceutical industry.There are some descriptions about the function of Epimedium brevicornu regulating blood glucose in many ancient books and traditional Chinese medicine books,which is one of the important biological functions of Epimedium brevicornu.Therefore,Epimedium brevicornu is frequently used in diabetes health care products in the folk,and is often used to relieve the symptoms of diabetes in clinic practice.The hypoglycemic effect of Epimedium brevicornu has attracted wide attention,and the research on the active ingredient is increasing.Literature research indicates that the hypoglycemic effect of Epimedium brevicornu may be related to the content of polysaccharide,glycoprotein etc.On this basis,the extraction,purification,characterization,antioxidation activity and hypoglycemic activity of the active ingredient of Epimedium brevicornu were studied in this paper.It is of great significance for the future development of the diabetes health food,the application of Epimedium brevicornu resources and the research on hypoglycemic activity.Ultrasonic/microwave synergistic extraction(UMSE)has been optimized to isolate polysaccharide from Epimedium brevicornu(EbLP).The optimum condition for UMSE was particle size 0.15×0.15 mm2,liquid-solid ratio 30 mL·g-1,extraction time 5 min.Full wavelength scanning test,Molish test,Fehling test and iodine-starch test were performed on EbLP,indicating that EbLP were non-starch polysaccharides.Three kinds of polysaccharides with different compositions were obtained by grading-alcoholic precipitation of EbLP,recorded as EbLP ?(The ethanol content is 30%,v/v),EbLP ?(The ethanol content is 30%,v/v)and EbLP ?(The ethanol content is 30%,v/v).Among them,EbLP ? had excellent scavenging activity on DPPH and ABTS free radicals,and the IC50 value were 0.42 mg·mL-1 and 0.54 mg·mL-1.EbLP ? can inhibit the activity of ?-amylase and ?-glucosidase,and the IC50 value were 24.0 mg mL-1 and 1.1 × 10-1 mg·mL-1,respectively.The diabetic model mice were treated with different doses of EbLP II for 21 d,the results as follows:1)model mice in high-dose group had a significantly lower food intake and water intake than those of the model mice in negative control(P<0.05),and the body weight was significantly higher than that of the model mice in negative control(P<0.05).2)The fasting blood glucose(FBG)and glycosylated serum protein(GSP)content in the model mice decreased rapidly(P<0.05),which was basically maintained at normal level.3)The swelling of liver and kidney in model mice was controlled,and the liver coefficient and renal coefficient were significantly decreased(P<0.05).4)The content of malondialdehyde(MDA)decreased significantly in the model mice,and the activity of glutathione peroxidase(GSH-PX)increased significantly(P<0.05).In conclusion,EbLP ? can adjust the FBG of diabetic model mice,and relieve the diabetic mellitus and complications.At the same time,it can improve the efficiency of scavenging free radical system,reduce the occurrence of lipid peroxidation,and protect and restore damaged tissues and organs.The glycoproteins of Epimedium brevicornu were obtained by soaking extraction method and salting-out method,then purified glycoproteins by dialysis,Seveg deprotein and column chromatography.The static adsorption and desorption tests of glycoproteins was performed on different macroporous adsorption resin(AB-8,D101,XDA-1 and X-5),and the content of glycoproteins was reflected in polysaccharides content and protein content,respectively.The results showed that the adsorption and desorption abilityies of D101 macroporous adsorption resin on glycoproteins of Epimedium brevicornu was prominent,and the adsorption saturation state could be reached within 3 hours.The results of dynamic adsorption and desorption tests were as follows:the best sample concentration was 0.075 mg·mL-1(in polysaccharide),and the best flow rate was 1.0 BV·h-1.The glycoprotein component ?,component ?,component ?,and component IV were obtained by using different polar solvent washing chromatography column.The SDS-polyacrylamide gel electrophoresis analysis showed that the component ? was belonged to glycoprotein.Further research reflected that the p? of component ? glycoprotein is 5.6,monosaccharide composition is:glucose,mouse plum,arabinose and galactose,and there is O-linked glycans in glycoproteins.Component II glycoprotein could effectively scavenging DPPH free radicals and ABTS free radicals,and the IC50 values were 1.49×10-2 mg·mL-1 and 3.80×10-2 mg·mL-1,respectively.The inhibitory effects of component ? glycoprotein on ?-amylase and a-glucosidase were dose-dependent,and the IC50 value were 1.15×10-1 mg·mL-1 and 8.33 ×10-2 mg mL-1,respectively.Above all,polysaccharides and glycoproteins of Epimedium brevicornu have prominent hypoglycemic activity,they has great effect on the treatment of diabetic complications,regulating blood glucose level of diabetic patients,improving the antioxidant capacity of the body and relieving symptoms of diabetes,and has the exploitation potential and development prospect.