Study On Isolation Purification And Immune Activity Of The Polysaccharides From Coptis Deltodiea C.y.cheng Et Hsiao.

Rhizoma Coptidis.is a traditional Chinese herbal medicine that is commonly used in treatment of heat-clearing and detoxification.Rhizoma Coptidis.is good at cleaning heat in middle jiao and their processed products are widely used in clinic.Wine Rhizoma Coptidis is good clear the fire of leader,Jiang Huanglian have anti-nausea effect,Yu Huanglian treat the disease of hot,Then there is a corresponding change on the pharmacological activity of the different processed products of Rhizoma Coptidis.Their main constituents are alkaloids,flavonoids and so on.Early studies focused on small molecules.In recent years,we found that single small molecules in on some efficacy can't replace the Rhizoma Coptidis.decoction.Therefore,it's necessary to the further research of pharmacodynamic material bases of Rhizoma Coptidis.Along with the further study of polysaccharides of Rhizoma Coptidis.,we found that it has antidiabetics,antioxidant and so forth.In this paper,we use of Radix Coptidis Deltoideae as raw materia to isolate and purify the crude polysaccharide.And study its physical and chemical properties and monosaccharide composition,then compare of the polysaccharide immune activity crude polysaccharide and processed polysaccharides.The main contents and results are as follows:1.Study the Radix Coptidis Deltoideae processed before and after that the polysaccharide chemical composition changes.Extract total polysaccharides processed before and after with the method of water extract-alcohol precipitation.The yield of polysaccharide of the original medicine is 2.08%,and the processed is 1.25%.Determinate polysaccharide,uronic acid and protein of total polysaccharides processed before and after by the means of sulfate-carbazole method and coomassie brilliant blue method.The crude content is 25.90%?2.59%respectively,and the processed is 33.27%?2.43%respectively.The protein content of the crude and the processed is 0.86%?0.82%respectively after using the Savag to eliminate protein.2.We apply Amberlite FPA90Cl-(Cl-)and Amberlite FPC3500-(H+)anion cation anion series resin column,DEAE Cellulose 52 to isolate and purify the crude polysaccharide.Then we got three kinds of polysaccharides C-1?C-2?C-3,and there were proved to be pure polysaccharide by HPLC.Their yields were 0.75%?0.525%?0.675%respectively,and molecular weights were 4.85×107Da,4.33×107Da,3.85×107Da.3.Applying the UV-VIS,IR,HPLC,and GC-MS to evaluate preliminary monosaccharide composition and structure of three polysaccharides.The results that PMP pre-column deriving HPLC analyzes show C-1 consists of ribose,glucuronic acid,galacturonic acid,glucose,fucose and their corresponding molar ratio was 1.2:1.9:4.1:8.2:1.C-2 consists of mannose,rhamnose,glucuronic acid,galacturonic acid,arabinose and their corresponding molar ratio was 2.7:4.3:8.1:8.3:1.C-3 consists of mannose,ribose,rhamnose,glucuronic acid,galacturonic acid,xylose and their corresponding molar ratio was 27.4:22.6:10.6:117.1:31.4:1.4.Research the immune activity of polysaccharide by char-coal clearance experiment,DNFB induced DTH,ConA and LPS induced spleen cell proliferation.The results shows different concentrations of total polysaccharides are able to change the value of carbon clearance index K and phagocytic index a value so DNFB-induced ear edema in mice DTH rate changes,polysaccharides exert immunomodulatory effects.After processing polysaccharides can even increase the value of carbon clearance index K and phagocytic index ? value,DNFB-induced ear swelling rate of DTH mice increased,with some immune-enhancing effect.The crude and the processed polysaccharide have synergistic inhibitive effect on spleen cell proliferative induced by ConA,LPS.The three kinds of homogeneous polysaccharide isolated all could inhibit spleen cell proliferative induced by ConA.In this paper we Study the Radix Coptidis Deltoideae prepared with the wine before and after that the polysaccharide composition and pharmacological activities changes.And we proved the biological active ingredients not just alkaloids that people have always considered while the polysaccharides also have good biological active.This reaserch laid a foundation not only for the development of medical value but also the future pharmacological activity research.Therefore,it has a important significance on Radix Coptidis Deltoideae polysaccharides structures' further in-depth study.