Study On The Separation,Structure Characterization And Absorption Of Anti-T2DM Polysaccharides In Radix Pseudostellariae

Pseudostellaria heterophylla is commonly used as a tonic agent in traditional Chinesemedicine and has the effects of invigorating qi and invigorating the spleen and nourishing the lung,from the dry root of Pax ex Pax et Hoffm,for spleen body tired,loss of appetite,weakness after illness,lack of qi and Yin,spontaneous sweating,dryness,dry cough and other symptoms.Diabetes is known in traditional Chinese medicine as the thirst disease,Qi and Yin double supplement,invigorating qi and invigorating the spleen is an important way of thirst disease in traditional Chinese medicine,so,Pseudostellaria heterophylla is widely used in the prescription of Chinese medicine for the treatment of thirst disease.In the early stage of the research group,the water extraction and alcohol precipitation method is used to classify the polysaccharides of Pseudostellaria heterophylla and to obtain different levels of Pseudostellaria polysaccharides.Based on the animal models of type 1 diabetes(T1DM)and type 2 diabetes(T2DM),the hypoglycemic study of different level ofPseudostellaria polysaccharides,The results showed that the level of Pseudostellaria polysaccharide on type I diabetes animal model only weak hypoglycemic effect and hypoglycemic effect on animal models of type Ⅱ diabetes mellitus significantly,the molecular weight of 50kDa-200kDa range of Pseudostellaria polysaccharides(PF40)hypoglycemic effect is particularly significant.Pseudostellaria polysaccharides(PF40)can improve insulin resistance and glucose tolerance in type Ⅱ diabetic animal models and improve insulin resistance.Pseudostellaria polysaccharides have hypoglycemic effect.At present,the study on the hypoglycemic effect of Pseudostellaria polysaccharides only stays on the level of crude polysaccharides of pseudostellaria heterophylla,and there is no in-depth study on its separation,purification and structure characterization.The different purity of pseudostellaria polysaccharides,different molecular weight of pseudostellaria polysaccharides and different structure of pseudostellaria polysaccharides and its hypoglycemic activity may have difference.So,in this experiment,water extraction and alcohol precipitation.DEAE-cellulose and dextran gels with different exclusion ranges were combined to separate and purify pseudostellaria polysaccharides,and homogenous pseudostellaria polysaccharides were obtained.Break through the level of crude polysaccharides,reached the level of homogenous polysaccharides.In this study,pseudostellaria polysaccharide PF40 were graded by different concentrations of ethanol.The isolated pseudostellaria polysaccharide PF40 was further isolated and purified to obtain homogenous pseudostellaria polysaccharides.The monosaccharide composition of the pseudostellaria polysaccharides was determined by the pre-column derivation method.The structure of the homogenous pseudostellaria polysaccharides was analyzed by using a variety of spectral analysis techniques,such as optical spectrometer,UV,IR,NMR,etc.,and the spatial structure of the polysaccharide was further studied.Caco-2 cells are a human cloned colon adenocarcinoma cell,whose physiological structure,function,and biochemical role are similar to human intestinal epithelial cells.In recent years,the Caco-2 cell model is an in vitro model for the study of drug small intestinal absorption at home and abroad.Pseudostellaria polysaccharide itself does not contain chromophores,This topic using fluorescein isothiocyanate(FITC)fluorescent labeling of pseudostellaria polysaccharide,it is easy to detect and trace the fluorescent groups,the Caco-2 cell model was used to simulate the absorption of homogenous pseudostellaria polysaccharides in human small intestinal epithelial cells,and the absorption pathway of pseudostellaria polysaccharides in vivo were explored.In this experiment,uniform homogenous pseudostellaria polysaccharides were obtained by separating and purifying pseudostellaria polysaccharides,Using modern pop technology to analyze its structure,the structure-activity relationship of homogenous pseudostellaria polysaccharides is more clearly defined.In addition,the in vitro model was used to simulate the absorption of small intestine to study the absorption pathway of homogenous pseudostellaria polysaccharides and lay a foundation for the development of Taizi ginseng hypoglycemic agents.