Extraction,Purification,Characterization And Antitumor Activity Of Polysaccharide From Roots Of Radix Tetrastigma

Tetrastigma hemsleyanum Diels et Gilg,belonging to the family Vitaceae,is a herbaceous perennial species native to China.As a popular folk medicine,the entire herb,especially its root was widely used for the treatment of fevers,children convulsions,owning to its anti-inflammatory,analgesic and antipyretic properties.Polysaccharides are polymeric carbohydrate structures,which have been widely investigated due to their strong biological activities.The polysaccharide from roots of Radix Tetrastigma(RTP)were extracted by water bathed method,and the yield of polysaccharide was determined by response surface methodology.Based on the single factor test,the optimum conditions were as follows: temperature 96?,water/liquid ratio 34:1 m L/g,extraction time 2 h.The yield of polysaccharide was(21.23 ± 0.77)%.After deprotein with sevag method,RTP-1?RTP-2?RTP-3 were successively purified by DEAE-52,respectively.Then they were further purified by Sephadex G-75?Sephadex G-200 and Sepharose CL-4B gel fltration chromatography,which named RTP-1-1?RTP-2-1?RTP-3-1,respectively.The purifed polysaccharide was designated as RTP-3-1,while its components,molecular weight,primary structure and domains were identified by high performance gel permeation chromatography(HPGPC),Gas Chromatography(GC),ultraviolet spectroscopy and infrared spectroscopy.The results showed that RTP-3-1 was high purity polysaccharides with a molecular weight of 1244.2 KDa,and it is mainly composed of 4 kinds of monosaccharides: arabinose,galacturonic acid,galactose,ructose,the proportion is 8.39%,7.18%,20.70%,63.70%,its had no obvious peaks of nucleic acid and protein,but had the characteristic absorption peak of acidic polysaccharide.1HNMR showed RTP-3-1 was an ?-type polysaccharide.RTP could significantly inhibit the proliferation of human gastric cancer SGC-7901 cells.The possible way is to induce cell apoptosis by decreasing the mitochondrial membrane potential.