Bl2基因与神经生长因子（NGF）抑制神经细胞凋亡的研究.docVIP

Bcl-2基因与神经生长因子(NGF)抑制神经细胞凋亡的研宄 韩贵和魏威顾军(浙江中医药大学附属中丙医结合医院骨科浙江杭州 310003) 】目的探讨原癌基因bcl-2与祌经生长因子(NGF)联合应用对抑制祌经细胞凋亡的协同作用。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、E、F六组，A组 为 bcl-2-PC12 细胞培养液中不加 H2O2 及 NGF (bcl-2-PC12 组)；B 组为 bcl-2-PC12 细胞培养液中加H2O2 (bcl-2-PC12+H2O2组)；C组为bcl-2-PC12细胞培养液中 加H2O2及NGF (bcl-2-PC12+ H2O2+NGF组)。D组为NC-PC12细胞培养液中不加 H2O2 及 NGF( NC-PC12 组);E 组为 NC-PC12 细胞培养液中加 H2O2( NC-PC12+ H2O2 组)；F组为NC-PC12细胞培养液中加H2O2及NGF (NC-PC12+H2O2+NGF组)。 皮用流式细胞仪检测各组细胞的凋亡率，釆用BCA(bicinchoninic acid)法检测bcl-2 基因表达蛋白浓度，数据进行统计学分析。结果A组细胞凋亡率较D组低(u=2.16, 0.02lt;Plt;0.05), B组较 E 组低(u=2.38，0.01lt;Plt;0.02)，C 组较 B 组低(u=2.27, 0.02lt;Plt;0.05)及较 F 组低(u=2.83,0.02lt;Plt;0.05),统计学分析有显著性差异。 A组bcl-2基因表达蛋白浓度高于D组(t=2.87, 0.005lt;Plt;0.002), B组高于 E 组(t=2.75, 0.01lt;Plt;0.05), C 组高于 B 组(t=2.16, 0.02lt;Plt;0.05)及 F组(t=2.23, 0.02lt;Plt;0.05)，统计学分析有显著性差异。结论bcl-2基因及 NGF均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，二 者联合应用时具有协同作用。 关键词】bcl-2基因祌经生长因子祌经细胞凋亡 】R39 】A 】2095-1752 (2013) 27-0056-03 Abaissement role of bcl-2 gene and NGF on the apoptosis of nerve cells HANGuihe,WEI Wei,GUJunl (Department of orthopaedics,The Affiliated Clinical Hospital of Integrated Traditional and Western Medicine of Zhe Jiang University of Traditional Chinese Medicine,Zhejiang Hangzhou,310003,China) Abstract】 Objective To approach synergia abaissement role of the proto-oncogene bcl-2 and NGF on the apoptosis of nerve cells.Methods PC12 cells were cultured to the logarithmic growth phase,slow virus plasmid carrying the bcl-2 gene and slow virus plasmid infected respectively PC12 cells by 100 MOl.Then they were divided into six groups(groupA,groupB,groupC ,groupD,groupE and groupF ). Group A:PC12 cells with slow virus plasmid carrying the bcl-2 gene.Group B: PC12 cells with slow virus plasmid carrying the bcl-2 gene, which were damaged by H2O2.Group C: PC12 cells with slow virus plasmid carryingthe bcl-2 gene, which were damaged by H2O2, were treated with NGF.Gr