| Dendrobium candidum is one of the traditional expensive medicines in our country, which has high pharmacological value. Dendrobium polysaccharides are the most import medicinal ingredients. In this paper, Dendrobium was selected as materials, and the polysaccharide was obtained from Dendrobium candidum, the polysaccharide DCP1 with component was collected through separation and purification, the component, IR spectrat of DCP1 were analyzed the antioxidant activity was studied in vivo and vitro, the antitumor activity of DCP1 on cell proliferation and apoptosis in tumor cell was studied in vivo and vitro, and mechanisms underlining above biological effects was explored. The above studies can provide some theoretical basis for application and exploration of Dendrobium candidum and some theory for the functional study of Dendrobium candidum polysaccharide. The main contents and results are shown as follows:(1)Among three extraction methods for polysaccharides from Dendrobium candidum, compound enzymes treatment method was selected finally for its excellent effect. After deproteinization and removed small molecular, the polysaccharide was purified by DEAE-52 cellulose ionic column. The result showed that the deionized water was better than NaCl solution of O.lmol/L. Number 5 to 11 cuvettes of eluent were collected for further study, after freeze-drying, we got the polysaccharide DC-1. DC-1 was subjected to futher purification on a Sephadex G-100 gel filration column, DCP1 was collected, the single symmetrical peak showed that DCP1 was pure.(2)The monosaccharide composition and structure of DCP1 was analyzed by HPLC and IR.HPLC analysis showed that DCP1 is mainly composed of mannose, rhamnose, glucose, galactose, fucose with molar of 1.748:0.973:1.241:1.017:0.561Infrared spectroscopy analysis indicated that DCP1 has typical characteristic absorption peaks. In combination with other characteristic absorption peak, it is determined that DCP1 was a type of pyrane polysaccharide of β-glucosidal bond with no uronic acid.(3)DPPH free radical scavenging, ABTS free radical scavenging, reducing power used to determine the antioxidant activity of DCP1.The results indicated that DCP1 has antioxidant activities in vitro, and the antioxidant activity depends on the concentration of DCP1, but the effect is less than Vc.(4)We studied the effect of DCP1on aging mice induced by D-galactose, activity of T-SOD, CAT and content of MDA in serum and liver were determined. The results showed that activity of T-SOD, CAT in mice which treatment were higher than aging model group, but compared with control group decreased; serum and liver MDA levels were higher than that of the control group, but compared with ging model group were decreased, indicated that DCP1 could significantly elevate the activity of T-SOD, CAT and decrease the MDA content in mice serum and liver.(5)Inhibition of DCP1 on HepG-2, H22 was determined with an MTT assay. The results showed that DCP1 could inhibit HepG-2, H22 tumor cells in different degrees; in the 0-640ug/mL range, with the increase of the concentration of DCP1 the inhibition rate gradually increased, action time from 36h to 48h, the inhibition rate increased a little; Cell apoptosis morphologic changes of HepG-2 were examined using AO/EB staining. The results showed that a series of morphology characteristics of apotosis on HepG-2 was by the decotion of DCP1.(6)The transplantable models of H22 in mice were established to investigate the inhibitory effects of DCP1. The main results indicated as follows:the decotion of DCP1 could inhabitation of solid on liver cancer H22 cancer, could improve the immunity of mice,the mumber of IL-2, TNF-a increased with ELISA assay, the solid tumor tissue pathology appear the apotosis features; DCP1 could increase life span in ascites H22. The decoction of DCP1 may induce apoptosis and enhance immunity to inhibit of the H22 tumor cell in vivo. |
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