| Object: To investigate whether the polysaccharide from jujube (JPS) alone or in combination with chemotherapeutic drug 2-methoxy-4-hydroxyl-5-acetylazonbenzene-4'-arsenic acid (R2) could anti-tumor in vivo and in vitro. Methods: (1) The dried jujube were extracted with hot water, the extract was concentrated under lower pressure and used ethyl alcohol to precipitate the crude polysaccharide. Then remove the fattiness, protein and pigment contents. The crude extracts were separated by anion-exchange chromatography on a DEAE-cellulose. (2) The anti-tumor activities of JPS and R2 were studied by cell culture in vitro. The inhibitory effects on HepG-2 cells were evaluated by MTT technique. Apoptosis was analyzed by cell proliferation, cell viability and morphological changes. Assay the apoptosis rate and cell phase of HepG-2 cells by using the flow cytometer. (3)The bearing tumor mouse model with Sarconma-180 was established for anti-tumor effect in vivo, then use JPS and R2 alone or united to treat the mice. After treatment, Weigh and kill the mice. Pick off the eyes and imbibe blood to measure WBC and PLT. Strip and weigh the neoplasm block heart, liver, kidney, spleen and thymus, calculate the inhibition rate of the anti-tumor, spleen index, thymus index and liver index. Use Pathological section to examine the injured status of the neoplasm and bowels; Use electronic microscope to observe the neoplasm cells. Results: (1)The coarse jujube polysaccharides were preliminarily purified by means of skellysolve G degrease, enzyme unite chloroacetic acid butanol to remove protein, H2O2 depigmentation, ethanol precipitation。Two large components, nominated JPS-N and JPS-A, were fractionated from the coarse polysaccharides by DEAE-cellulose column chromatography, eluted by distilled water and 0~1mol/L sodium caproate. (2) The growth of HepG-2 cells which were treated by JPS or JPS combination with R2 could be significantly inhibited and have dose-time-dependent effects. JPS and R2 may induce apoptosis when be used singly or together. But There is on obviously increase apoptosis when use the two drugs (3) The synergism that JPS improve the effect of R2 through the bearing tumor model of Sarcoma-180. The variations of the body weight, thymus index, spleen index, white blood cell (WBC) and blood platelet (PLT) amount of tumor bearing mice show that JPS has significant effect on all of above index and JPS can confront the leucopenia and thrombocytopenia caused by R2. Pathological section shows that both JPS and R2 could damnify the neoplasm and only R2 could damnify the liver and kidney. Under electron microscope, cell shrinkage, chromatin concentration and formed irregularly shaped crescents at the nuclear edges were observed. Finally, fragmentation of cell into membrane-wrapped apoptotic bodies was appeared. The results of the synergism experiment show that JPS could significantly improve the effect of R2 and decrease the toxicity of R2 in vivo. Conclusion: The results in this present study demonstrated that JPS and R2 both had anti-tumor effect in vivo and vitro. These effects were dose-time-dependent and might be related its immune-regulation and induction of tumor cell apoptosis. JPS and R2 have synergism anti-tumor activity to some extent in vivo and in vitro. |
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