| Radix pseudostellariae polysaccharide(RPP)is a major effective ingredient of radix pseudostellariae.It has been shown to be immunomodulatory,anti-stress,anti-inflammatory,fatigue resistance and so on.It shows that selenizing modfication is a widely used method to enhance the biological activities of Radix pseudostellariae polysaccharide,it is of great significance to modify the radix Pseudostellariae polysaccharide with selenium.This article selected the radix pseudostellariae polysaccharide(RPP)as the research object.To investigate the effects of selenizing radix Pseudostellariae polysaccharide(sRPPs)on the activity of cellular immunity,RPP was extracted by water decoction and ethanol precipitation,purified through eliminating protein by Sevage method and column chromatography of DEAE-52,then selenizingly modified by nitric acid-sodium selenite method according to L9(34)orthogonal design of three-factors,the usage amount of sodium selenite,reaction temperature and reaction time,at three level to obtain nine selenizing RPPs,sRPP1?sRPP9.To observed the effects on immune adjustment at different doses and different types of radix pseudostellariae polysaccharides(RPP)and selenzing radix Pseudostellariae polysaccharide(sRPPs),we selected the RAW264.7 macrophage as cellular model.Otherwise,to observed the effects on immune adjustment at different doses and different types of RPP and sRPPs,we selected the immunity damaged mice which caused by eyclophosphamide(CY)as test objects.The test results are as follows:1 The effects on RAW264.7 macrophages proliferation in vitro were compared by MTT assay.The results showed that sRPPs could significantly promote macrophages proliferation,sRPP2,sRPP3,sRPP8 and sRPP9 presented stronger efficacy.To investigate the effect of sRPPs on the immune activity of the RAW264.7 macrophage,added the sRPP2,sRPP3,sRPP8 and sRPP9 on macrophage,respectively;The yield of NO was measured by Griess method;The secretion levels of TNF-a and IL-6 in RAW264.7 macrophages was determined by ELISA method.The results showed that these four sRPPs could promote the relative survival rate,phagocytic uptake,NO production,TNF-a and IL-6 secretion of RAW264.7 cells.These results indicated that selenizing modification could significantly enhance the immune-enhancing activity of RPP.2 The determination of sRPPs on the immune function of the immune injury model in mice induced by cyclophosphamide.220 Balb/c mice were randomly divided into 11 groups,9 treatment groups were injected sRPP2,sRPP3 and RPP(0.2 mL)1,2,3 mg/mL via subcutaneous injection,control group and CY model group was given saline,for 3 days.4th days,treatment group and CY model group were injected of cyclophosphamide(CY)via intraperitoneal injection(80 mg/kg/d)for 3 d.In the last injection after 24 h,draw four in each group with carbon clearance test in mice macrophage phagocytosis index,and the rest of the mice were weighed and then cervical dislocation executed after enucleating eyeball blood,detected the levels of cytokine and immunoglobulin,immune organ index,spleen and thymus index.The results showed that the sRPPs and RPP can promote the immunity damaged mice of phagocytotic function of mononuclear phagocytic system,receded the influence of cyclophosphamide to the immune organs,and receded the influence of IgG and IgM content in serum which caused by cyclophosphamide,meanwhile decreased the influence of production the IL-6,TNF-a,which caused by cyclophosphamide in immunity damaged mice.Conclusion:According to above-mentioned results to make a comparison of modified polysaccharide to non-modified polysaccharides,it can be seen that the activities of sRPPs are significantly stronger than RPP in enhancing phagocytosis,immune organ index secreting NO,and cytokines of peritoneal macrophages,immunoglobuiilt index.This indicates that selenizing modification could improve the immune-enhancing activity of polysaccharides. |
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