Extraction Of Polysaccharides From Epimedium Leaf,Purification And Its Prevention Effects On Alcoholic Liver Injury

Epimedium Koreanum Nakai which is classed as a plant of the Berberaceae family and known as Herba Epimedii is a type of traditional Chinese medicinal material with a long history of medicinal use.Studies found that it has rich medicinal effects,such as anti-inflammatory,anti-aging,and hypolipidemic effects.In recent years,studies have shown that Epimedium leaf resources are effective in protecting the liver.At present,most of the researches on Epimedium is focused on flavonoids and alkaloids.However,there are few reports on the physiological activity of Epimedium polysaccharide,and its mechanism of action is not yet clear.Therefore,polysaccharides from the leaves of Epimedium were extracted and purified,the basic structures of the polysaccharides were explored,and the liver protection effects and mechanisms of the epimedium leaves polysaccharides were revealled.The main findings are as follows:1.Ultrasonic water extraction method was first used to extract polysaccharides from Epimedium brevifolia leaves,and orthogonal experiment was used to optimize the extraction conditions of polysaccharides.The optimal extraction conditions were determined as the following:the ratio of material to liquid is 1:25,the extraction temperature is 70?,and the extraction time is 50 minutes.The extraction rate of Epimedium polysaccharide obtained by using the best extraction conditions was 5.98%.A homogeneous polysaccharide component with a purity of 82.03% was obtained by using the DEAF-Sepharose fast flow anion exchange column and the Sephadex G-100 dextran gel column to separate and purify the polysaccharide.The molecular weight was determined to be 31.2 k Da by high-performance gel permeation chromatography.The structure of Epimedium polysaccharide was analyzed by infrared spectroscopy,and the result showed that the Epimedium polysaccharide has a pyranose structure.The monosaccharide compositions hydrolyzed by Epimedium leaf polysaccharide were investigated by means of acid hydrolysis,derivatization analysis and high performance liquid chromatography.Experimental results showed that the compositions were arabinose(2.25): galactose(1.12): glucose(0.74): glucuronic acid(1.00): rhamnose(1.15): mannose(0.32).2.Establish a mice liver injury model induced by alcohol,and explore the mechanism of Epimedium leaf polysaccharides in preventing alcoholic liver injury.The test was divided into 5 groups: blank group,model group,low,medium and high doses of Epimedium(10,20,40 mg / kg).The results showed that the medium and high dose groups of Epimedium polysaccharide could reduce the content of AST or ALT in serum,the content of MDA and TG in blood and liver compared with the model group,and increase the activities of SOD and GPX and the content of GSH in blood and liver of model group.When acute alcohol was ingested,the medium and high doses of Epimedium polysaccharide could increase the SOD activity and GSH content of the liver mitochondria of the model group,and reduce the MDA content at 10 h;All dosage groups can increase the GSH content in the liver of the model group and reduce the MDA content at 15 h and 20 h;Pathological sections suggested that the medium and high dose groups of Epimedium improved necrotic and denatured liver tissues,and the high dose group had the best performance.It indicated that Epimedium polysaccharide can protect liver tissue.3.The combination of oleicacid(OA)and ethanol was used to induce Hep G2 cells to establish an alcoholic fatty liver(AFLD)cell model,which was divided into groups of blank group,BSA group,model group(with 1 mmol / L of OA and 56% Ethanol),and three groups of Epimedium polysaccharides in different concentrations(200,400,800 ?g / m L).The experimental results showed that all dosage groups inhibited the increase of AST,ALT and TG in the injured cells,effectively reduced MDA content in cells and increase SOD activity;Meanwhile,the group of Epimedium polysaccharide in 400 ?g / m L significantly increased the activity of ADH,reduced the accumulation of ethanol in liver cells,which had a significant effect on improving the aleoholic fatty live disease function.