The Regulation And Mechanism Of Selenized Codonopsis Polysaccharide On The Function Of Bone Marrow Dendritic Cells

Selenide polysaccharide has many pharmacological effects,such as enhancing immune function,anti-oxidation,anti-tumor,hypoglycemic and hypidemia,and its biological activity is higher than that of selenium and polysaccharide.In this experiment,crude polysaccharide was extracted from Codonopsis pilosula by water extraction and alcohol precipitation method,the protein was removed by TCA method,and DEAE-52,G-200 separation and purification,then the polysaccharides of Codonopsis pilosula were selenized by nitric acid-sodium selenite method,and the effects and mechanism of codonopsis pilosula polysaccharides on dendritic cell function before and after selenification were studied.The purpose of this study was to improve the biological activity of codonopsis pilosula polysaccharide and provide theoretical basis for the preparation of polysaccharide traditional Chinese medicine immune enhancer.Results show that:1.The yield of codonopsis pilosula polysaccharide and codonopsis pilosula polysaccharide without protein were 10.92% and 4.2%,respectively;The sugar contents were 43.36% and 57.40%,respectively;DEAE-52 with H2 O elution had the highest sugar yield and sugar content were 2.54% and 83.81%,respectively;by further separation and purification of codonopsis pilosula polysaccharide by G-200,the sugar yield and sugar content were 52.36% and 98.86%,respectively.2.The selenium content of codonopsis pilosula polysaccharide after selenization was12.11 mg/g.The phenotype was observed by scanning electron microscope,fourier transform infrared spectroscopy suggests characteristic absorption peaks,the molecular weight was determined by high performance gel permeation chromatography,the monosaccharide composition was analyzed by PMP pre-column derivatization and the C and H structures were identified by nuclear magnetic resonance spectroscopy.The results showed that selenized codonopsis pilosula polysaccharide had smooth,compact sheet structure with Se=O and Se-O-C characteristic absorption peaks,and molecular weight less than 1.0 KD,and mainly contained Glu,Gal,Ara,Man and Glc N and were α and βstructured.3.Flow cytometry was used to detect the effects of s CPPS on the phagocytic activity of immature BMDCs and the phenotype of mature BMDCs.It was found that s CPPS significantly enhanced the phagocytic activity of immature BMDCs,significantly increase the expression of phenotype MHCⅡ,CD86,CD80 and CD40 of mature BMDCs,significantly increase the contents of IFN-γ,IL-12 and IL-1β in cell supernatant and significantly enhance the ability of BMDCs to stimulate the proliferation and antigen presentation of allolymphocytes.4.Western Blot was used to detect the effect of s CPPS on TLR signaling pathway.It was found that s CPPS could significantly increase the expression of My D88,NF-κB P65 and NF-κB P-P65 in My D88/NF-κB signaling pathway.Conclusion:1.Selenized codonopsis polysaccharide was prepared by nitric acid-sodium selenite method,and its structure was identified,indicating that the selenization modification of codonopsis polysaccharide was successful.2.Compared with CPPS,s CPPS can significantly enhance the function of immature and differentiated mature BMDCs,indicating that selenized Codonopsis pilosula polysaccharide can enhance the immune function of the body.3.The regulation mechanism of s CPPS on BMDCs function is realized by activating My D88/NF-κB signaling pathway.