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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。
AIM: To investigate the protective effect of ginseng protein (GP) on neurons induced by H2O2 in mice. Methods: Neurons were isolated and purified from neonate neonatal rat cerebral cortex. Cell morphology was observed by inverted fluorescence microscope. Cell senescence was detected by β-galactosidase kit. Cell viability was detected by MTT assay. Apoptosis was detected by Hoechst 33342 / PI double staining And necrosis, fluorescent quantitative RT-PCR assay apoptosis gene Bcl-2, Bax mRNA expression. Results: Hoechst33342 and β-galactosidase positive staining were found in H2O2-induced morphological changes such as shortening or disappearance of axons, rounding and shrinking of cell bodies, varying sizes, nuclear pyknosis and nuclear chromatin accumulation. Ginseng protein can improve cell morphology, increase cell survival, reduce apoptosis and necrosis, reduce cell aging rate. CONCLUSION: Ginseng protein has a protective effect on neurons induced by H2O2. Its mechanism is related to promoting the expression of Bcl-2 mRNA and inhibiting the expression of Bax mRNA.