人参蛋白对过氧化氢致神经元损伤的保护作用

目的:采用H₂O₂诱导小鼠神经元损伤,研究人参蛋白(GP)对神经元的保护作用。方法:从新生乳鼠大脑皮层中分离纯化神经元,通过倒置荧光显微镜观察细胞形态,β-半乳糖苷酶试剂盒检测细胞衰老,MTT法检测细胞存活,Hoechst 33342/PI双染色法检测细胞凋亡与坏死,荧光定量RT-PCR实验检测凋亡基因Bcl-2、Bax mRNA 表达。结果:H₂O₂致细胞出现轴突缩短或消失,胞体变圆缩小,大小不等,核固缩,核染色质聚集等形态学变化,细胞存活率降低,Hoechst33342及β-半乳糖苷酶染色阳性;人参蛋白可改善细胞形态,增加细胞存活率,减少细胞凋亡与坏死,降低细胞衰老率。结论:人参蛋白对H₂O₂致神经元损伤具有保护作用,其机制与促进Bcl-2 mRNA表达,抑制Bax mRNA表达有关。

Neuroprotective effects of ginseng protein on neurons against damages induced by hydrogen peroxide

OBJECTIVE To investigate neuroprotective effects of ginseng protein (GP) on mice neuron damages induced by hydrogen peroxide (H₂O₂).METHODS Primary neurons, isolated and purified from cerebral cortex of suckling mouse, were observed for cell morphology by using inverted fluorescence microscope. Cell senescence was detected by β-galactosidase assay kit, cell survival by MTT colorimetry, apoptosis and necrosis of cells by Hoechst 33342/PI double staining method, and Bcl-2 and Bax mRNA expression by fluorescent quantitative RT-PCR.RESULTS After treatment of H₂O₂, axons shortened or disappeared, cell bodies became round, with reduced and unequal size. Karyopyknosis and aggregation of nuclear chromatin happened, cell survival rate decreased, and Hoechst 33342 staining and β-galactosidase staining showed positive results. Besides, ginseng protein could improve cell morphologies, increase cell survival rate, decrease cell apoptosis and necrosis, and reduce rate of cell senescence.CONCLUSION Ginseng protein has neuroprotective effects on neurons against damages induced by H₂O₂, which may be associated with induction of a pro-apoptotic gene Bcl-2 and inhibition of anti-apoptotic gene Bax mRNA expression in neurons.