| Objective Platycodon grandiflorum as raw material,extraction polysaccharide form Platycodon grandiflorum,separation and purification obtained pure polysaccharide.Then its structure was selenide modified,and studies on process conditions,then investigated the biological activity and compared.Methods Using assisted extraction technology of polysaccharide in Platycodon grandiflorum by supercritical CO2,water extraction and ethyl alcohol method obtained polysaccharide,the optimum conditions of extraction technology is based on the yield index and content of polysaccharide;Optimization of preparation technology by orthogonal design the process conditions of supercritical CO2.First,the crude Platycodon grandiflorum polysaccharide are used DEAE-52 column initial separation and purification,then,using Sephadex G-75 column was purified secondary.Phenol-sulfuric acid method was used to detect the polysaccharide content of Platycodon grandiflorum polysaccharide;coomassie blue staining method was used to detect the protein content of Platycodon grandiflorum polysaccharide.Platycodon grandiflorum polysaccharide as raw material,adding sodium selenite performed Platycodon grandiflorum polysaccharide structure modification,generated organic selenium compounds(selenide Platycodon grandiflorum polysaccharide).Selenium content of organic selenium compounds was detected by HPLC to determine the optimum process,yield and selenium of Platycodon grandiflorum polysaccharide content for indicators.Platycodon grandiflorum polysaccharide,selenium Platycodon grandiflorum polysaccharide by autoxidation pyrogallic acid scavenging superoxide anion radical,scavenging effect of the hydroxyl radicals by Salicylic acid method,scavenging effect of DPPH radical,compared with Vc to prove Oxidation resistance.The nitro phenyl-alpha-D-pyran glycosidase as raw material,determination of nitrophenol by UV-vis.Platycodon grandiflorum polysaccharide containing selenium and inhibitory effects of Polysaccharide on aglycosidase and determine the inhibitory type.48 mices were divided into six groups(normal,control model,positive control,platycodon grandiflorum selenide polysaccharide low,medium and high dose group).After a week of normal feeding,continuous intragastric administration 14 days and 1 time everyday,the high dose group was administered 2.0 g·kg-1,middle dose group was 1.0 g·kg-1,low dose group was 0.5 g· kg-1 At the same time the positive bifendate control group was 1.0 g· kg-1,normal group and model control group was given the same volume saline.After 1 hour of the seventh and last administration,the groups of mice were injected with a solution of 0.2% CCl4 olive oil 10 m L·kg-1 except normal group.After 24 hours,obtain blood from eye globe and separated serum.Dislodge liver and put into normal saline.They were examined including ALT,AST,ALP,GGT,ALB,GLB,TP levels in serum and GSH,MDA levels in liver tissues.In addition,pathologic changes were examined by HE staining.Results The optimum conditions of supercritical CO2 fluid extraction: the temperature is 45 ?,extraction pressure is 25 MPa,ethanol concentration of 70%,the time is 1.5 h.When the use of DEAE-52 chromatography separation,one obvious peak of polysaccharide is isolated by distilled water,the quality of is 0.1342 g,yield to 28.21 %,one obvious overlapping peaks of polysaccharide and protein is isolated by 0.5mol/L Na HCO3,only protein peak but no polysaccharide is isolated by 1.0mol/L of Na HCO3.When the use of Sephadex G-75 gel filtration chromatogram purity,one platycodon grandiflorum polysaccharide obtained by tripledistilled water.Yield to polysaccharide content for 79 %,the quality of polysaccharide is 3.8 mg.The optimum process for selenide Platycodon grandiflorum polysaccharide were reaction temperature of 60 ? and time of 5 hours,the 6 % catalyst concentration,a 60 % ultrasonic frequency.Under these conditions,the average selenium content of 2.282 mg/g,and average extraction rate of 15.66 %.ID50 of Platycodon grandiflorum selenium polysaccharide value could not be measured in urgent poison experiment,and less toxicity.For different radicals,comparative Platycodon polysaccharide,Platycodon selenium polysaccharide and Vc scavenging.Oxidation resistance results expressed with IC50 alue concentration.Superoxide anion radical scavenging ability respectively were 1.021 mg/m L,0.512 mg/m L and 0.042 mg/m L;hydroxyl radical scavenging ability respectively were 1.269 mg/m L,0.458 mg/m L and 0.596 mg/m L;DPPH radical scavenging ability respectively were0.312 mg/m L,0.230 mg/m L and 0.156 mg/m L.At the same concentration,Vc of total capacity reduction is highest,followed by selenium polysaccharide,the lowest is polysaccharide.The largest inhibition rate of Platycodon grandiflorum polysaccharide,Platycodon grandiflorum selenium polysaccharide and acarbose is 70.07 %,79.78 % and 94.21 % respectively,the IC50 is 1.10 mg/m L,0.82 mg/m L and 0.41 mg/m L,respectively.The type of Platycodon grandiflorum polysaccharide is Competitive inhibitory,and Platycodon grandiflorum selenium polysaccharide is the competition and competitive inhibition of hybrid.Platycodon grandiflorum polysaccharide on acute liver injury model mice can be reduced to varying degrees of ALT,AST in serum and elevated GSH levels in liver tissue,which can improve CCl4-induced pathological tissue damage.Conclusions After supercritical CO2 extraction technology assisted extraction,the yield of polysaccharide and polysaccharide content is high,the extraction process is simple,avoid the use of chemical methods,reducing the chemical pollution of polysaccharide;Process of polysaccharide selenium are simple,and easy to control,content of selenium is high,and antioxidant of Platycodon grandiflorum selenium polysaccharide are stronger than Platycodon grandiflorum polysaccharide.Selenium polysaccharide and polysaccharide had inhibitory effect on the hypoglycemic activity of a-glycosides.This study confirms Platycodon grandiflorum polysaccharide on CCl4-induced acute liver injury has a protective effect,the mechanism is likely to be reached by scavenging free radicals and prevent liver damage. |
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