人参蛋白协同H_2O_2诱导SH-SY5Y细胞氧化损伤

Oxidative injury in SH-SY5Y cells induced synergistically by Ginseng protein and H_2O_2

目的:探讨人参蛋白(GP)协同H2O2诱导人神经母细胞瘤(SH-SY5Y)氧化应激损伤的作用,并筛选二者的最佳联合浓度。方法:首先采用不同浓度H2O2诱导SH-SY5Y细胞氧化损伤,然后采用不同浓度GP联合200μmol·L-1 H2O2诱导SH-SY5Y细胞氧化损伤;采用倒置荧光显微镜观察细胞形态,MTT法检测细胞存活,Hoechst 33342染色法检测细胞凋亡。结果:SH-SY5Y细胞经GP-H2O2作用后,细胞数量减少,轴突缩短或消失,胞体变圆、缩小,大小不等;细胞存活率降低;Heochst 33342染色呈现高蓝光;GP 60mg·L-1+H2O2200μmol·L-1为联合诱导氧化应激损伤的最佳浓度。结论:GP有协同H2O2诱导SH-SY5Y细胞氧化损伤的作用,抑制细胞增殖、促进细胞凋亡是其可能的作用机制。

OBJECTIVE To investigate oxidative stress injury induced by ginseng protein（GP）and H2O2 in human neuroblastoma SH-SY5 Ycells,and screen optimal combined concentration of GP and H2O2.METHODS SH-SY5 Ycells were induced firstly by different concentrations of H2O2,and then oxidative stress injury was induced synergistically by different concentrations of GP and H2O2（200μmol·L-1）.Cell morphology was observed by inverted fluorescence microscope,cell survival determined by MTT colorimetry,and cell apoptosis detected by using Hoechst 33342 staining method.RESULTS Number of SH-SY5 Ycells was decreased,axons of cells were shortened or even disappeared,cells displayed round and shrunken cell bodies in different sizes,and cell survival rate was decreased.Besides,Hoechst 33342 showed positive staining.Optimal combined concentration was GP（60mg·L^-1）＋H2O2（200μmol·L^-1）for GP and H2O2-induced oxidative stress injury.CONCLUSION GP has synergistic effects on H2O2-induced oxidative injury in SH-SY5 Ycells,and its action mechanism may be inhibition of cell proliferation and promotion of apoptosis.