1.Separation and purification of total alkaloids from Evodia rutaecarpa using 732 cation exchange resin and investigations on its anti-breast cancer bioactivity
Shixue CHEN ; Yuanyong YAO ; Zhongying LU ; Bangcheng TANG
Chinese Journal of Biochemical Pharmaceutics 2015;(9):15-18
Objective To optimize conditions for extraction of alkaloids from Evodia rutaecarpa by means of 732 cation exchange resin and performe its anti-breast cancer bioactivity.Methods The optimum processing route on extraction of alkaloids from Evodia rutaecarpa was investigated by means of 732 cation exchange resin.The performance of 732 cation exchange resin was compared with tranditonal process ( aqueous extraction-ethanol precipitation in extraction-purification process).The alkaloid reagent-potassium mercuric iodide test and MTT test were performed on the crudes.Results Compared with traditional purification process, it was much better to use 732 cation exchange resin approach for extraction of alkaloids from Evodia rutaecarp with high yield(16.81%) The best eluent should be saturated brine with excellent purification.No obvious correlations were found between the toxin of breast cancer cell and concentration of total alkaloids.When the concentration of total alkaloids was 50μmol/mL, cellular survival rate was 68%afterward 24 h.When the concentration of total alkaloids comes to 100 μmol/mL and 150 μmol/mL, cellular survival rates were slightly decreased by 66% and 60%.Conclusion 732 cation exchange resin performes much higher than traditional process in purification of total alkaloids extracted from Evodia rutaecarpa.Simultaneously, the extracted total alkaloids showe remarkable inhibition in breast cancer cell.
2.Effect of inhibition of TGF-β1 receptors on lymphocytes during infection of Echi-nococcus granulosus
Shuanghong YIN ; Junbo ZHANG ; Xiaolin CHEN ; Fangjie XU ; Jun HOU ; Xiangwei WU ; Xueling CHEN ; Yuanyong YAO ; Mingming XING ; Wencai YANG
Chinese Journal of Immunology 2015;(5):607-612
Objective:To investigate the effects of TGF-β1 on T lymphocytes of BALB/c mice infected with Echinococcus granulosus( E.granulosus ) in vitro.Methods: The inhibitor group:the spleen cells of BALB/c mouse were co-cultured with E.granulosus and SB525334.The control group:the spleen cells of BALB/c mouse were co-cultured with E.granulosus and PBS.The blank group:the spleen cells of BALB/c mouse were co-cultured with RPMI-1640 medium and SB525334.The lymphocytes were collected at 48 h post-infection.The T lymphocyte subsets, the number of CD4+CD25+T cells, the number of NK cells, and the expression of NKG2D receptor were detected by flow cytometry.The NK cell activity was determined with the lactate dehydrogenase leakage assay(LDH).Results:The inhibition the TGF-β1 receptors resulted in the increase of in the number of CD4+T cells,the decrease in the number of CD8+T cells,the increase of in the ratio CD4+/CD8+T cells,the decrease of in the number of CD4+CD25+T cells,the increase in the expression of the NKG2D receptors,the increase in the lysis rate of Yac-1 cells by NK cells,and a positive cor-relation between the expression of activity receptor NKG2D and killing activity of NK, which were mediated by E.granulosus.Conclusion: The inhibition of TGF-β1 receptors can enhance the immune response of T lymphocytes against E.granulosus infection in vitro.