Maca Polysaccharide Promotes CD4~+T Proliferation And Research Its Adjuvant Effect On Chemotherapy

Objective:To investigate the therapeutic effect of Maca polysaccharide assisted with 5-fluorouracil(5-FU)on transplanted lung cancer in mice,and to analyze its possible mechanism,and to verify it through in vitro and in vivo experiments to build experimental basis for new adjuvant chemotherapy drugs.Method:(1)The effect of Maca polysaccharide on proliferation and apoptosis of Lewis cells was detected by MTT assay and Annexin V/7-AAD assy.(2)The lung cancer bearing mice model was constructed by subcutaneous injection of Lewis cells.The tumor-bearing mice were intraperitoneally injected with5-FU,and Maca polysaccharides were given by intragastric administration.Observe the growth of subcutaneous tumor in mice,observe the size of the tumor and measure its volume;measure and calculate the thymus index,spleen index and tumor inhibition rate of lung cancer bearing mice.(3)To detect the proportion and number of CD4~+T cells,CD8~+T cells in spleen and lymph node,the proportion and number of B cells,DCs,mononuclear macrophages,MDSCs,CTL cells,Th1 cells,Th2 cells and Treg cells in spleen by flow cytometry(FCM);The content of IFN-γand IL-4 in serum was detected by ELISA.The expression of p53,Bax and Bcl-2 in tumor tissue was detected by Western-blot.(4)MACS was used to isolate wild-type mouse CD4~+T cells,and after treatment with Maca polysaccharide and 5-FU,CFSE assay was used to detect the change of CD4~+T cells proliferation.Results:(1)In vitro treatment of Lewis cells with Maca polysaccharide showed that Maca polysaccharide had no obvious inhibitory effect on the proliferation of Lewis cells,but Maca polysaccharide could promote the early apoptosis of Lewis cells.(2)Compared with the 5-FU control group,Maca polysaccharide-assisted 5-FU treatment group had slower tumor growth and decreased tumor volume.(3)Compared with the 5-FU control group,the Maca polysaccharide-assisted5-FU treatment group had reduced tumor weight in mice.Compared with the control group,the tumor weight of mice in the 5-FU control group was significantly reduced(p<0.005).Compared with the 5-FU control group,500 mg/kg of Maca polysaccharide assisted 5-FU treatment group and 1000 mg/kg,the tumor weight of Maca polysaccharide-assisted 5-FU treatment group was significantly reduced(p<0.05),and the average tumor inhibition rate was increased.Compared with the control group,the Maca polysaccharide-assisted 5-FU treatment group both increased spleen index and thymus index.(4)The results of FCM showed that the proportion and number of CD4~+T cells and CD8~+T cells in spleen and lymph nodes were significantly decreased after 5-FU treatment,but The proportion and number of CD4~+T cells and CD8~+T cells were significantly increased(p<0.05),and the effect was more pronounced when the Maca polysaccharide concentration was 1000 mg/kg.After treatment with 5-FU,the number of B cells was significantly decreased in the spleen,and the number of B cells treated with 1000 mg/kg of Maca polysaccharide assisted with 5-FU was significantly increased(p<0.05).Compared with the 5-FU control group,the use of 1000 mg/kg maca polysaccharides assisted in the 5-FU treatment slightly increased the proportion of Th1 cells in the spleen.Compared with the blank control group,the proportion of CTL cells decreased after 5-FU treatment,1000 mg/kg maca polysaccharide assisted5-FU treatment increased the proportion of CTL cells significantly(p<0.05).There was no statistically significant change in the proportion and number of Treg cells,Th2cells,DCs,MDSCs,and mononuclear macrophages in spleen and Th2 cells in lymph nodes after maca polysaccharide-assisted 5-FU treatment.(5)Compared with the control group,Bax and p53 were significantly up-regulated in the 5-FU control group,and Bcl-2 expression was decreased.Compared to the 5-FU control group,Maca polysaccharide-assisted 5-FU treatment group showed up-regulation of Bax and p53.(6)Compared with the blank control group,the expression level of IFN-γin serum of the 5-FU control group mice was decreased,but after the treatment with Maca polysaccharide,the expression level of IFN-γwas slightly increased,but there was no statistical significance;Compared with the control group,the serum level of IL-4 in the 5-FU control group mice was decreased.After the treatment with Maca polysaccharide,the expression level of IL-4 did not change significantly.(7)In vitro,5-FU was added to the proliferation system of CD4~+T cells.After 72hours of culture,proliferation of CD4~+T cells was significantly inhibited;different concentrations of maca polysaccharide were added to the proliferation system of CD4~+T cells.After 72 hours,the proliferation of CD4~+T cells was enhanced.When both Maca polysaccharide and 5-FU were added simultaneously,the proliferation of CD4~+T cells suppressed by 5-FU could be partially restored after 72 hours of culture.Conclusion:(1)Maca polysaccharide combined with 5-FU can inhibit tumor growth in lung cancer transplanted mice.(2)Maca polysaccharide can inhibit Lewis cells by promoting the apoptosis of Lewis cells.(3)In the tumor-bearing mice,Maca polysaccharides can promote the apoptosis of tumor cells and restore the damage of 5-FU immune organs.Maca polysaccharide can increase the proportion of CD4~+T cells,CD8~+T cells,B cells,Th1 cells,and CTL cells in vivo.(4)Maca polysaccharide can promote the proliferation of CD4~+T,and it can restore the proliferation of CD4~+T when 5-FU inhibits the proliferation of CD4~+T.