Study On Antioxidant Effect Of Pumpkin Polysaccharide And Ginsenoside Or Rutin In Vitro

In this paper,the crude polysaccharide of pumpkin was used as raw material for separation and purification.The molecular weight of pumpkin polysaccharide was determined by GPC method.The composition of monosaccharide was determined by high performance liquid chromatography(HPLC),as well as far infrared spectrum,ultraviolet full-wavelength scanning,scanning electron microscope image,etc.In the experimental content,the anti-oxidation effect of pumpkin polysaccharide and ginsenoside Rg1 and rutin was studied by scavenging free radicals and other antioxidant experiments.The pumpkin polysaccharide and ginsenoside Rg1 were combined for in vitro cell antioxidant test,in order to be used in functional foods.The application and product development provide relevant reference.The main contents of this paper are as follows:(1)The crude polysaccharide of pumpkin was deproteinized by Sevage method.The polysaccharide content of pumpkin crude polysaccharide was determined by phenol sulfuric acid method to be 20.81%.The crude polysaccharide of pumpkin was passed through DEAE-52 anion exchange chromatography column and collected 0.3 mol/L.The fraction eluted with Na Cl was dialyzed and lyophilized.The next step was purified by Sephadex G-200 gel chromatography column to obtain a purified pumpkin polysaccharide having a polysaccharide content of 92.39%.(2)The molecular weight of pumpkin polysaccharide was determined by GPC method.The molecular weight of pumpkin polysaccharide was about 6.076×102 kD.The far-infrared spectroscopy and monosaccharide composition analysis showed that the purified pumpkin polysaccharide was acidic heteropolysaccharide,and the monosaccharide composition was high-performance liquid.Chromatographic analysis,the pumpkin polysaccharide is composed of mannose,ribose,glucosamine,glucuronic acid,galacturonic acid,glucose,xylose,galactose,and aminogalactose;the pumpkin polysaccharide is found by ultraviolet full-wavelength scanning.No protein characteristic absorption peak at 280 nm,the purification effect is better.The purified pumpkin polysaccharide and ginsenoside Rg1 and their mixtures were observed by scanning electron microscopy.The crude polysaccharide was pleated in a spherical shape.The crude polysaccharide except the protein was irregularly shaped with pores,and the purified polysaccharide was in the form of flakes.Ginsenoside Rg1 is a smooth and irregular crystal.It can be seen from the scanning electron microscope image that the mixed pumpkin polysaccharide and ginsenoside Rg1 are in a smooth sphericalstate,which is different from the electron microscope image of pumpkin polysaccharide and ginsenoside Rg1 in a single state.(3)The free radical scavenging and lipid peroxidation experiments in vitro were divided into 6 groups,namely positive control BHA,pumpkin polysaccharide,rutin,ginsenoside Rg1,pumpkin polysaccharide and rutin compound,pumpkin polysaccharide and ginsenoside Rg1 compound,The control and the sample were subjected to gradient concentration treatment to determine the ability to scavenge DPPH free radicals,hydroxyl radicals and lipid peroxidation.The results showed that the compound of pumpkin polysaccharide and ginsenoside Rg1 was stronger than the single group in the determination of DPPH free radical scavenging ability.The compound of pumpkin polysaccharide and ginsenoside Rg1 was free to DPPH in the experiment by the addition method.The removal of the base has a certain synergistic effect;while the ability to scavenge hydroxyl radicals and lipid peroxidation,the synergistic effect of pumpkin polysaccharide and ginsenoside Rg1 is not obvious(P>0.05),and pumpkin polysaccharide and ginseng were found during the experiment.The antioxidant capacity of the saponin Rg1 complex was significantly stronger than that of the single group(P<0.05).The pumpkin polysaccharide and rutin compound were analyzed by the addition method,and it was found that it did not have obvious synergistic effect(P>0.05).The antioxidant capacity of pumpkin and rutin compound was stronger than that of pumpkin polysaccharide group,lower than the rutin group.(4)The newborn SPF mice were fed for one week,and the liver of the mice was taken to obtain hepatocytes.The mouse hepatocytes were induced by H2O2 to establish a model of oxidative stress damage.In the experiment,the mice were divided into 5 groups,namely normal group,model group,pumpkin polysaccharide group,ginsenoside Rg1 group and pumpkin polysaccharide and ginsenoside Rg1.The experimental concentration gradients were set to 0.333 mg/mL and 0.667 mg/ respectively.mL and 1 mg/mL.MDA content,GSH-Px activity,SOD activity and CAT activity were measured,respectively.The results showed that the single group of 0.667 mg/mL and 1 mg/mL pumpkin polysaccharide and ginsenoside Rg1 and their combination group acted on oxidative stress injured cells,and the combined group had significant repair effect on oxidative stress injured cells(P<0.05),can significantly improve the activity of cells,alleviate the damage caused by oxidation,enhance the antioxidant capacity of cells,and have a dose-effect relationship.When the concentration was 0.333 mg/mL,the single group and the combined group acted on oxidative stress-damaged cells,although the effect was not as good as the medium-highdose group,but it could also improve the effect.