Extraction And Biological Activity Of Crude Polysaccharide From Polygonum Oiliinerve (Nakai) Ohwi

Polygonum cillinerve(Nakai) Ohwi,one of the most valued traditional Chinese medicines,is a member of "Taibai Qiyao" and first year's growth liane,commonly found in hillside,forest,bosk, and lawn in the Taibai mountains and many northwestern provinces in China.As a drug,it can be traced back to tu jing ben cao.Whole plant may be taken for medicine.It has functions of nourishing the kidney,nourishing the liver,and anti-aging,immuno-modulating,anticancer,antibacterial, antivirus,and anti-microorganism activities.It is commonly used in China for over 2000 years for the treatment of quinsy,enteritis,gastritis diarrhea,nephritis dropsy,allantois infection and so on.In this paper,polysaccharide in Polygonum cillinerve(Nakai) Ohwi is studied,about which there was no reports as yet.There are two parts in this thesis:Part one:review1.A survey of study on Polygonum cillinerve(Nakai) OhwiIt evolved chemical component and pharmacological activity of P.cillinerve(Nakai) Ohwi.2.A survey of study on polysaccharideThere are four faces in the research advancements of polysaccharide:research advance of polysaccharides;sources of polysaccharides;a review on the situation of the pharmacological activities of polysaccharides;and the application and the development direction of the studies on polysaccharides.3.Research advance of antioxidationCharacteristics of free radical;relation between oxygen free radical and diseases,aging; antioxidation system in vivo;the system of evaluation of antioxidation in vitro;and antioxidant activity and applications of traditional chinese medicine.Part two:research papers In this part,four departments are involved:extraction,isolation and purification of the polysaccharides in Polygonum cillinerve(Nakai) Ohwi;Anti-oxidation of polysaccharides in vitro; Anti-oxidation of polysaccharides;anti-tumor activity in vitro of polysaccharides.First,extraction,isolation and purification of the crude polysaccharides in Polygonum cillinerve (Nakai) Ohwi(PCCP).The sample was defatted with petroleum ether,remoned the essential oils with steam distillation.The water-soluble polysaccharide was extracted by boiling water from the sediment,and then was precipitated by 80%(v/v) of ethanol.The crude polysaccharides in Polygonum cillinerve(Nakai) Ohwi obtained is red.The compounds with small molecular weight were removed with dialyser and a part-pure product of polysaccharide was obtained,and the total contents of polysaccharides were determined by the method of sulphuric acid-phenol.PCCP is decolorized by DEAE-cellulose chromatography.The results showed extraction yield of polysaccharide by boiling water was about 5.1%;the total contents of polysaccharides in PCCP was about 91.8%.The DEAE-cellulose chromatography could purify polysaccharides preliminarilySecond,hydroxyl radical scavenging,super-oxide anion radical scavenging,DPPH.radical scavenging and lipid per-oxidation inhibiting were introduced to assay the antioxidant activity of PCCP in vitro.The results showed:hydroxyl radical,super-oxide anion radical,DPPH.radical was scavenged by 35.1%,40.7%and 38.3%when the concentration of PCCP comes to 8.48 mg/mL; Lipid per-oxidation inhibition rate of PCCP was 62.0%at 8.48 mg/mL.The results indicated that PCCP has antioxidant activities in vitro.Third,PCCP were studied for their anti-oxidation and immune responses activity in mice.The in vivo investigation demonstrated that administration of PCCP by gavage is able to overcome cyclophosphamide-induced immunosuppression and significantly raised total antioxidant capacity, catalase activity,superoxidase dismutase activity,glutathione peroxidase level.It also raises liver, spleen and thymus indices,and decreases malondialdehyde in a dose-dependent manner in mice.The results indicated that the PCCP,possessing pronounced free radical scavenging and antioxidant activities,could play an important role in prevention of oxidative damage in immunological system.Lastly,the anti-tumor activity of PCCP was evaluated by MTT colorimetric assay with HepG-2. HepG-2 cell is a cell line derived from HCC.This investigation showed that PCCP could inhibite HepG-2 cell proliferation in vitro.HepG-2 inhibition rate of PCCP was 42.6%when the concentration of PCCP comes to 277.78μg/mL,which indicated that PCCP has anti-tumor effects in vitro.The polysaccharides of Polygonum Cillinerve(Nakai) Ohwi was firstly studied in this paper.It was preliminarily proved that PCCP has antioxidation and antitumor activities.The biological activities have made a foundation for the further study and application of the polysaccharides of Polygonum Cillinerve(Nakai) Ohwi.