| S.rugosoammulata Falow is a fungus of the genus Stropharia of the Strophariaceae family,which is also one of the top ten varieties in the mushroom trading market in the world.It is grown in many developing countries.S.rugosoammulata is delicious.It has bright color and considerable yield.The studies have shown that the fruity body of S.rugosoammulata is rich in a variety of beneficial minerals,proteins,vitamins,polysaccharides and other nutrients,which has certain edible value and medicinal value as well as development potential.It is of great significance to study the structural characteristics,physicochemical properties and activity evaluation of polysaccharide biomolecules for the exploitation and utilization of S.rugosoammulata resources.S.rugosoammulata fruiting body has a variety of biological activities,can effectively improve the body immunity.Therefore,the development of mushroom polysaccharides has positive effect on the development of edible mushroom industry.In this paper,the polysaccharide of S.rugosoammulata was used as the main object of study.The single factor combined with orthogonal test was used to comprehensively evaluate the antioxidant and immunomodulatory effects of the macroscopus macroscopus model extracted from the three-phase method and the in vivo immunosuppression experiment.In order to provide raw materials and theoretical basis for the development of functional food or natural medicine.The main research results are as follows:(1)With the extraction rate of polysaccharide from the fruity body of S.rugosoammulata(SRP)by three-phase method as the target,the control parameters were sulfuric acid mass fraction,the volume ratio of sample liquid to t-butanol,time and temperature.Orthogonal test was used to determine the optimal extraction process of polysaccharides from S.rugosoammulata as follows:mass fraction of sulfuric acid20%,volume ratio of sample liquid to t-butanol 1:1.5,temperature 35°C.Under the above conditions,the yield of SRP was 6.85%.The monosaccharide composition showed that SRP was composed of 8 monosaccharides,the molecular weight showed that SRP was 27.52 k Da,and the FT-IR showed that SRP had the infrared absorption peak of sugar.XRD result showed that SRP contains amorphous structure.SEM result showed that SRP exists in fragmentary form and has depression after magnification.(2)In vitro antioxidant experiment results showed that SRP had good scavenging ability of DPPH,hydroxyl radical,ABTS~+free radicals and good reducing power.The results of CCK-8 experiment showed that SRP concentration in the range of 25-200μg/m L could promote cell proliferation without cytotoxicity.The results of neutral red phagocytosis experiment and scratch experiment showed that in the range of 25-50μg/m L,SRP can significantly promote phagocytosis and scratch healing of macrophage RAW264.7,and also promote the expression of cytokine mRNA and induce autophagy by up-regulating the expression of key proteins in the TLR4/NF-κB signaling pathway.These results indicated that SRP had certain immune activity in vitro.(3)The immunosuppressive model of mice was established by intraperitoneal injection of cyclophosphamide,and the immune-enhancing effect of polysaccharide from S.rugosoammulata was evaluated.The results of immunomodulatory activity in vivo showed that SRP could significantly relieve the immunofunction of mice induced by cyclophosphamide,and could significantly improve the body weight and spleen index of mice.The contents of Ig G,Ig A,TNF-α,IL-6 and IL-1βin serum of mice were significantly increased,and the value of CD4~+/CD8~+in splenic lymphocytes was significantly increased,indicating that SRP could effectively relieve immunosuppression. |
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