The Research Of The Homogeneous Polysaccharide From Rumex Gmelini Turcz On Anti-Tumor And Immune Activity In Vitro And Component Analysis

Decades of scientific experiments confirmed that the polysaccharide has extensive and complex biology activities,because it has no cytotoxicity,and little side effects to organisms.There are thorough researches no matter from the polysaccharide extraction ,purification process,composition structure analysis and biological activities.Most plant polysaccharide can inhibit or kill tumor cells,without damage to normal cells,it can inhibit tumor growth,activate immune cells,improve the immune function of organism[I].This experiment mainly study the extraction,separation and purification of the polysaccharide from Rumex gmelini Turcz,Screening its anti-tumor immune activity components,analyzing its compositions,deepening the level of studying polysaccharide from Rumex gmelini Turcz.Objective This topic is the polysaccharide from Rumex gmelini Turcz as the research object,it has a series of experiments of extracting,separation and purification of polysaccharide from Rumex gmelini Turcz,and get the homogeneous polysaccharide. Combining with the previous studies that the anti-tumor effects of the polysaccharide from Rumex gmelini Turcz,exploring its antitumor immune effects of homogeneous polysaccharide,analyzing its compositions,deepening the level of studying polysaccharide from Rumex gmelini Turcz.Providing the basis of the study of the medicinal plant development and the comprehensive utilization of the resources of Chinese medicine.Methods Using hot water to get Rumex gmelini Turcz root water extraction,using water extraction-alcohol precipitation method by 95% ethyl alcohol to get crude polysaccharide from Rumex gmelini Turcz,clearing off the protein,pigment and other works such as filtering out impurities,content determinations with phenol vitriolic colorimetry.Using DEAE cellulose column chromatography and Sephadex G-100 gel column chromatography to further purify polysaccharide,and get two homogeneous polysaccharide RKA,RKB,using HPLC-ELSD method measuring two polysaccharide purity.CCK-8 method is used to screening out the two homogeneous polysaccharide inhibition of tumor department (liver cancerHepG-2,lung cancerA549, colon cancerSW1116, bladder cancerT24, breast cancerMCF-7) and the content that can inhibite turner cells. Inverted microscope and flow cytometry instrument are used to observing the cell morphology,under the effect of RKA macrophages to secrete cytokines quantity change.Experiments have established that RKA has obvious inhibitory effect of tumor cells in vitro,it has certain influence on the immune.Adopts full hydrolysis sugar alcohol acetate derived gas phase-mass spectrometry are uniform polysaccharide RKA composition analysis and high performance liquid gel column chromatography method to calculate the molecular weight distribution.Results Phenol-sulfuric acid method for determination of the content of polysaccharide from Rumex gmelini Turcz in a standard curve to calculate the polysaccharide content is 78.84%.DEAE cellulose column chromatography elution curve shows the highest water elution,the study of homogeneous polysaccharide from Rumex gmelini Turcz of arteries and veins,Sephadex G-100 gel column chromatography shows elution two relatively single component,elution curve,HPLC-ELSD detection show the single peak symmetry,showed that both groups are divided into homogeneous polysaccharide from Rumex gmelini Turcz, namedRKA and RKB.RKA present certain inhibition on HepG 2,A549,MCF-7,SW1116, among them the HepG-2,the MCF-7 has significant inhibitory effect.RKB for selected tumor inhibition is weak.RKA much way to activate macrophages, promote the secretion of cytokines, immune enhancement effect.lt is composed of arabinose,xylose,mannose and glucose four kinds of monosaccharide composition,mole ratio is 1.79:1:4.46:1.2.Main ingredients molecular mass of 1.14 ×104 Da.Conclusion This experiment get two homogeneous polysaccharide from Rumex gmelini Turcz by extraction and purification,and identifid to be two relatively single component. RKA has obvious inhibiting tumor effect,it may be related to its promoteing macrophage factor secretion and enhancing immune function.