Study on the Effect of Astragalus Polysaccharide on the Proliferation of Human Neuroblastoma SH-SY5Y Cells / 中国药房

ABSTRACT

OBJECTIVE:To investigate the inhibitory effect of astragalus polysaccharide(APS)on the proliferation of human neuroblastoma SH-SY5Y cells. METHODS:After the cells were cultured with 0(blank control),25,50 and 100 mg/ml APS for 6,12 and 24 h,MTT method was used to determine cell viability and calculate inhibition rate. Following cell cultured with 0 (blank control),25,50 and 100 mg/ml APS for 24 h,Hoechst 33258 fluorescent staining was performed,and then cell nucleus morphology was observed under the fluorescence microscope;flow cytometer was used to detect the distribution of cell cycles and apoptosis;western blot was employed to determine the expression of extracellular regulated protein kinases (ERK) 1/2 protein in cells. Enzyme linked immunosorbent assay (ELISA) was conducted to determine the contents of interleukin 2 (IL-2),IL-6 and IL-12 in the cells. RESULTS:Compared to the blank control,those cultured with 100 mg/ml APS for 6 h,50 and 100 mg/ml APS for 12 h and 25,50 and 100 mg/ml APS for 24 h demonstrated higher inhibition rate. After the cells were cultured with 50 and 100 mg/ml APS for 24 h,those in G0/G1 phase increased and those in G2/M and S phases decreased,and the contents of IL-2 and IL-6 increased. After cells were cultured with 25,50 and 100 mg/ml APS for 24 h,the apoptosis rate was higher,densely hyperchromat-ic fragments in cell nuclei and apoptotic bodies appeared,the phosphorylation level of ERK1/2 protein in the cells was lower,and the content of IL-12 was higher. There was statistically significance (P<0.01 or P<0.05). CONCLUSIONS:APS can inhibit the proliferation of SH-SY5Y cells by arresting cell cycle and inducing cell apoptosis through a mechanism which may be correlated to the decrease in the phosphorylation of ERK1/2 and increase in cytokine.