Screening Of Active Components For Lycium Barbarum Polysaccharides To Repair Chemical Liver Injury

"Compendium of Materia Medica"records:Lycium barbarum L.has the function of nourishing the liver and treating liver and kidney yin deficiency.Modern research shows that Lycium barbarum polysaccharides have significant repair function on chemical liver injury and drug-induced liver injury.Lycium barbarum polysaccharides are the main effective components of liver damage and repair.However,as the molecular weight distribution of Lycium barbarum polysaccharides is widely distributed,the effective polysaccharide components for the repair of chemical liver damage have not been reported,which leads to the unclear base of its active substance,which restricts its application in health food,special medical use food and medicine.In this paper,the main main line of the study is to trace the material basis of Lycium barbarum polysaccharides in the repair of chemical liver injury.The separation and purification of Lycium barbarum polysaccharides are carried out by hierarchical alcohol precipitation or molecular exclusion chromatography,and the activity tracking of each component separated by the animal model experiment or chemical activity evaluation method is tracked.The Lycium barbarum is studied systematically.The material basis of sugar repair for chemical liver injury is to provide reference for the development of health food,special medical use food and drugs with Lycium barbarum polysaccharide for the repair of chemical liver injury.The main contents and results are as follows:1.Study on the biological activity of Lycium barbarum polysaccharides in repairing chemical liver injury.A model of CCl₄ liver injury was established,and the effect of LBP on liver injury in rats was explored through the study of general toxicity,liver histopathological changes,and serum liver function related indexes（serum protein concentration and serum related enzyme activity）in rats.The study showed that 100 mg·kg^-1and 200 mg·kg^-1of Lycium barbarum polysaccharide group could reverse the abnormal changes of serum ALB and TP caused by CCl₄,and significantly increase the activity of ALT,AST and gamma-GT.At the same time,compared with the liver protection tablets and the silybin group,the serum ALB,GLB,and ALB of the rats of the Lycium barbarum polysaccharide group of 100 mg·kg^-11 and 200mg·kg^-1were all different from the two.There was no statistical significance（p>0.05）.At the same time,there was no significant difference in serum ALT,AST,AKP,gamma-GT activity and TBIL content between the two groups of high dose Lycium barbarum polysaccharide group,respectively（p>0.05）.It showed that the high dose Lycium barbarum polysaccharide group was in the reverse of the abnormal changes of serum protein and the activity of liver function related enzymes in CCl₄.The curative effect of the liver protection tablets and the silybin is the same.The results suggest that 100 mg·kg^-11 and 200 mg·kg^-11 have significant effects on the repair of chemical liver damage,and the effect is similar to that of the positive control 900 mg·kg^-11 liver protection tablets and 30 mg·kg^-11 silybin.2.Screening of active fractions from Lycium barbarum polysaccharides in repairing chemical liver injury.LBP1,LBP2,LBP3 and LBP4 were obtained by fractionation of alcohol,and the activity of protective chemical liver injury of 4Lycium barbarum polysaccharides was systematically evaluated through the investigation of DPPH free radical scavenging ability,superoxide anion radical scavenging ability,total antioxidant capacity and hydroxyl radical scavenging ability.The study found that all 4 Lycium barbarum polysaccharides have the ability to scavenge DPPH free radicals,scavenging superoxide anion radicals and the ability to scavenge hydroxyl radicals.The 4 Lycium polysaccharides are the best in eliminating the effect of various free radicals,and the second is LBP3.Thus,it can be concluded that the ability of protecting the protective chemical liver damage should also be LBP2.The composition is best.This was in accordance with the order of the content of Lycium barbarum polysaccharides and the content of aluronic acid in the 4components,which showed that the protective activity of Lycium barbarum polysaccharides was positively related to the content of polysaccharide and the content of uronic acid.The ability of scavenging DPPH free radicals,superoxide anion radicals and hydroxyl radicals by 4 components shows that the ability to scavenge hydroxyl radicals is the strongest.It suggests that Lycium barbarum polysaccharide may be a way to repair chemical liver damage or protect the liver by removing hydroxyl radical.3.The basic research of Lycium barbarum polysaccharide in repairing chemical liver injury.Considering that Lycium barbarum polysaccharide is a kind of complex polysaccharide with glycopeptide structure,we studied whether or not protein was removed.LBP was separated and analyzed by HPLC-DAD-ELSD.Among the 3peaks obtained,the peak of 1 and 2 was a glycopeptide component,and the peak of No.3 was a polysaccharide component.The DPPH free radicals were scavenged by HPLC on line.The results showed that the effect of glycopeptide was best,so the protein was not considered in the subsequent purification.LBP2 was decolorized and purified by DEAE-52 column chromatography.In the elution curve,3 peaks were eluted,respectively,corresponding to wash off,0.1 mol·L^-11 NaCl elution and 0.3mol·L^-11 NaCl elution.Among them,LBP I was neutral polysaccharide,LBP II and LBP III were acidic polysaccharides and acid LBP II<LBP III.From the protein elution curve,LBP I almost did not contain protein,so the content of the purified polysaccharide reached 89.4%and the other two components were the complex of polysaccharide and protein.The free radical scavenging activity of LBP and LBP I^LBP III was screened and the results of LBP II scavenging were the best,indicating that LBP II was the active component of LBP,which may be related to the amount of hydrogen supply and the number of active groups,such as hydroxyl and amino groups.Sephadex G-100 column chromatography was used to purify LBP II.The elution curves showed 1 peaks and the peak was symmetrical.LBP II-1 was identified as homogeneous polysaccharide by HPLC,providing a basis for further structural studies.