Separation And Purification Of Japanese Metaplexis Polysaccharide And Protection Of Japanese Metaplexis Polysaccharide On Liver Injury In Mice Induced By CCl₄

ObjectiveTo study the Japanese Metaplexis Polysaccharide separation andpurification methods and preliminary study on hepatoprotective effect.MethodsUse Ⅱtype ZTC1+1natural clarify agent to refine the extractedJapanese Metaplexis polysaccharide compound to get crude polysaccharide,Through the hollow fiber ultrafiltration impurity removal of small molecules andscreening of Sephadex column specification，using DEAE-52cellulose column(Diethylaminoethyl-cellulose DE-52column)(2.6x50cm) purified crudepolysaccharide,Sephadex G-100gel chromatography column furtherpurification Japanese Metaplexis polysaccharide; With phenol-sulfuric acidmethod tracking and detecting Japanese Metaplexis polysaccharide contents;Using DEAE-52cellulose column and Agarose gel electrophoresis to purityidentification. To establish the CCl4mouse model of acute liver injury inducedby CCl4,experimented on the liver injury model of mice，determine the ALTactivity and AST activity of serum by automatic biochemical analyzer. ResultsNatural clarifying agent of ZTCl+l purification optimum Metaplexis totalsugar was A, B component of the adding amount of liquid volume respectively1%,2%, concentrated liquid for20mL/g, temperature70℃, heating time was1h/1h; The crude polysaccharides by DEAE-Cellulose-52and SephadexG-100gel column chromatography for further purification by two mainpolysaccharide fractions: JMP1-A, JMP3-B. Two kinds of polysaccharidesshowed single peak by Sephadex G-100gel column chromatography, agarosegel electrophoresis showed only one spots.Increase of Metaplexis japonicapolysaccharide dose group could against carbon tetrachloride induced acuteliver injury in mice AST and ALT, that of Metaplexis japonica polysaccharidescan cell membrane lipid peroxidation caused by against CCl4, protect the livercells and subcellular damage.ConclusionsApplication of ZTC1+1Ⅱnatural clarifier purification of Metaplexisjaponica polysaccharide, do not need to adjust the Asclepiadaceaepolysaccharides pH, only need to select the A, according to the nature of thepending liquid B component addition, purification of temperature,concentration and purification of time, can get good clarified effect, it isconvenient to use, and removal of protein and polysaccharide retention effectis good. Asclepiadaceae polysaccharide can cell membrane lipidperoxidation caused by against CCl4, protect the liver cells and subcellulardamage.