Studies On Antioxidant Activities Of Polysaccharide-Selenizing Polysaccharide Compounds

Selenium polysaccharide is one of organic selenium compounds.It was easily absorbed and utilized by the organism and possessed duple and higher biological activity of polysaccharide and selenium,such as enhancing immunity,anti-oxidantion,anti-cancer,anti-metal poisoning and so on.Because of the yield of selenium polysaccharide and the high cost,the prevention and cure of animal diseases mainly by polysaccharide compound.In present study,firstly,three polysaccharides and three selenizing polysaccharides were prepared,then 18 compounds were composed with seven polysaccharides and three selenizing polysaccharides with stronger antioxidant activities according to carbohydrate content at 9:1 ratio of polysaccharide to selenizing polysaccharide.Secondly,the best three compounds of thier antioxidant activities were screened out by radical scavenging test,then the antioxidant activities of three compounds were compared through the test of ability to resist oxidative damage of Chicken embryo liver cells and the test in vivo and the best compound CAP-sLBP was screened out.The purpose of this study is to validate the antioxidant effect of polysaccharide-selenizing polysaccharide compound,screen the best compound with antioxidant activities,provide the materials for developing antioxidantors.The tests are divided into four parts as follows:Experiment 1 Preparation of polysaccharides and their selenide polysaccharides Lycium barbarum polysaccharide(LBP),Atractylodes macrocephala polysaccharide(AMP)and Chinese angelica polysaccharide(CAP)were extracted by water extraction and ethanol precipitation method,eliminated protein by trichloroacetic,permeate through DEAE-52 sephadex column and freeze drying,the purified LBP,AMP and CAP were obtained.The sLBP,sAMP,sCAP were prepared by HNO3-NaSeO3 method,which the optimization of selenizing modification were described as previously mentioned.Yields and contents of polysaccharides were determined.The results show that the yields of three crude polysaccharides were from 8.26%to 15.23%,the yields of three selenizing polysaccharides were from 29.13%to 39.87%,the contents of polysaccharides of three polysaccharides and three selenizing polysaccharides were from 28.35%to 61.30%.Experiment 2 Comparison of their scavenging power of free radical of polysaccharide-selenizing polysaccharide compounds Based on our previous researches 18 compounds were composed with Lycium barbarum polysaccharide(LBP),Atractylodes macrocephala polysaccharide(AMP),Chinese angelica polysaccharide(CAP),Codonopsis pilosula polysaccharide(CPP),Garlic polysaccharide(GP),Lily polysaccharide(LP),Schisandra chinensis polysaccharide(SCP),selenizing LBP(sLBP),selenizing AMP(sAMP),and selenizing CAP(sCAP)with stronger antioxidant activities according to carbohydrate content at 9:1 ratio of polysaccharide to selenizing polysaccharide,and then diluted them with distilled water at five concentrations to determine their scavenging power of DPPH radical,hydroxyl radical and ABTS radical.The result showed that all compounds have different ability to scavenge three radicals,the clearance and dose were positively correlated.The top three order of average DPPH radical clearance rate were LBP-sAMP?CAP-sLBP and LBP-sCAP,the top three order of average hydroxyl radical clearance rate were LBP-sAMP?CAP-sAMP and SCP-sLBP,the top three order of average ABTS radical clearance rate were LBP-sAMP?CAP-sLBP and LBP-sCAP.In conclusion,the antioxidant activities of LBP-sAMP?CAP-sLBP and CAP-sAMP were better.Experiment 3 The effect of the ability to resist oxidative damage of CEL of polysaccharide-selenizing polysaccharide compounds Compared the effect of the ability to resist oxidative damage of Chicken embryo liver cells(CEL)of the three polysaccharide-selenizing polysaccharide compound(CAP-sLBP and LBP-sAMP and CAP-sAMP).At first testing the safe concentrations of three compound groups and VC group with MTT assay,then choose 3 different kinds of the safe concentration scope of the drug to join CEL training system,24 hours later adding H2O2 to attack the CEL for an hour.Determining the activity of CEL with MTT assay(the value of A570),testing the content of ROS in CEL,the changes of GSH-Px and SOD activity with the kit.The result showed that the cellular A570 value in H2O2 group was lowest,ROS contents were the highest,the activities of GSH-Px and SOD were the lowest,which has significant difference with cell control group.The cellular A570 value in 3 compound groups and VC group were higher or significantly higher than H2O2 group,ROS contents were lower or significantly lower than H2O2 group,the activities of GSH-Px and SOD were higher or significantly higher than H2O2 group,high concentrations were better.3 compounds each at 5 concentrations have the stronger antioxidant activity than LBP-sAMP and CAP-sAMP with same concentration.The result indicated that 3 compounds all have the stronger antioxidant activity,they can resist to CEL vitality decline,ROS contents increased and antioxidant activities decline caused by H2O2,and enhance ability to resist oxidative damage,the activity of CAP-sLBP is the strongest.Experiment 4 the effect of chicken antioxidant function of polysaccharide-selenizing polysaccharide compounds Compared the effect of chicken antioxidant function of the three polysaccharide-selenizing polysaccharide compounds(CAP-sLBP,LBP-sAMP and CA-sAMP).Two hundred and seventy 14-day-old chicks were divided randomly into 9 groups with thirty chicks each.The chickens except blank control(BC)group were vaccinated with Newcastle disease IV vaccine,repeated vaccination at 28 days old.At the same time as the every vaccination,each chick in six polysaccharide groups were intramuscularly injected respectively two kinds of the three polysaccharide-selenizing polysaccharide compounds 0.5 ml,in vaccination control and blank control group,with equal volume of physiological saline,once a day for three successive days.Collecting blood from wing veins on 7,14,21 and 28 days after the first vaccination,separating the serum and determinating the contents of CAT?SOD?GSH-Px and MDA.The result showed that the activity of CAT?SOD and GSH-Px of six polysaccharide groups at every time points were higher or significantly higher than those of the control group and the value of MDA of six polysaccharide groups at every time points were lower or significantly lower than that of the control group.The activities of antioxidant enzyme in CAP-sLBP group were the best while the value of MDA was the lowest.These results indicated that the three compounds all have the different antioxidant activities to improve the Chicken antioxidant capacity and the CAP-sLBP possessed the strongest efficacy.