Bcl-2基因抑制神经细胞凋亡的实验研究

Inhibiting Role of Bcl-2 Gene in the Apoptosis of Nerve Cells

目的:观察原癌基因bcl-2对神经细胞凋亡的抑制作用。方法:培养PC12细胞至对数生长期,用100感染复数(multiplicity of infection,MOI)的携带bcl-2基因的慢病毒质粒及未携带bcl-2基因的慢病毒质粒感染PC12细胞。再将其分为A、B、C、D四组,A组为携带bcl-2基因慢病毒质粒的PC12细胞(bcl-2-PC12细胞),B组为未携带bcl-2基因慢病毒质粒PC12细胞(NC-PC12细胞),C组为bcl-2慢病毒质粒PC12细胞加H2O(2bcl-2-PC12细胞+H2O2),D组为NC慢病毒质粒PC12细胞加H2O(2NC-PC12细胞+H2O2)。应用流式细胞仪检测细胞的凋亡率,采用BCA(bicincho ninic acid)法检测bcl-2基因表达蛋白。结果:A组细胞凋亡率低于B组(P<0.05),C组细胞凋亡率低于D组(P<0.05);A组bcl-2基因蛋白浓度高于B组(P<0.05),C组bcl-2基因蛋白浓度高于D组(P<0.05)。结论:bcl-2基因对正常神经细胞的凋亡有抑制作用,能够增强神经细胞的抗损伤能力。

Objective： To observe the inhibiting role of the proto-oncogene bcl-2 in the apoptosis of nerve cells. Methods： PC12 cells were cultured to the logarithmic growth phase and then slow virus plasmid carrying the bel-2 gene and slow virus plasmid infected PC12 cells by 100 multiplicity of infection, respectively. The PC12 cells infected were then divided into 4 groups. Group A： PC12 cells with slow virus plasmid carrying the bcl-2 gene; group B： PC12 cells with slow virus plasmid; group C： PC12 ceils with slow virus plasmid carrying the bcl-2 gene, which were damaged by H202; group D： PC12 cells with slow virus plasmid, which were damaged by H202. The rate of apoptosis was detected by flow cytometry. The proteinum concentration of bcl-2 gene ex- pression was detected by using bicinchoninic acid method. Results： The apoptosis rate of group A was lower than that of group B and the protein concentration of bcl-2 gene expression of group A was higher than that of group B, both of which were significantly different between the 2 gruups（P〈O.05）. The apoptosis rate of group C was lower than that of group D and the protein concentration of bcl-2 gene expression of group C was higher than that of group D, both of which significantly differed between the 2 groups（P〈0.05）. Conclusion： Bel-2 gene can inhibit apoptosis of normal nerve cells and can enhance resistance ability of nerve cell to be damaged.