CAJ | 학술논문

目的探讨原癌基因bcl-2与神经生长因子（NGF）联合应用对抑制神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）。应用流式细胞仪检测各组细胞的凋亡率，采用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=2.16，0.02<P<0.05), B组较E组低（u=2.38，0.01<P<0.02），C 组较B组低(u=2.27，0.02<P<0.05)及较F组低(u=2.83，0.02<P<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=2.87，0.005<P<0.002）， B组高于E组（t=2.75，0.01<P<0.05），C组高于B组（t=2.16，0.02<P<0.05）及F组（t=2.23，0.02<P<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。
목적탐토원암기인bcl-2여신경생장인자（NGF）연합응용대억제신경세포조망적협동작용。방법배양PC12세포지대수생장기，용100감염복수（multiplicity of infection，MOI）적휴대bcl-2기인적만병독질립급미휴대bcl-2기인적만병독질립감염PC12세포。재장기분위A、B、C、D、E、F륙조，A조위bcl-2-PC12세포배양액중불가H2O2급NGF（bcl-2-PC12조）；B조위bcl-2-PC12세포배양액중가H2O2（bcl-2-PC12+ H2O2조）；C조위bcl-2-PC12세포배양액중가H2O2급NGF（bcl-2-PC12+ H2O2+NGF조）。D조위NC-PC12세포배양액중불가H2O2급NGF（NC-PC12조）；E조위NC-PC12세포배양액중가H2O2（NC-PC12+H2O2조）；F조위NC-PC12세포배양액중가H2O2급NGF（NC-PC12+ H2O2+NGF조）。응용류식세포의검측각조세포적조망솔，채용BCA(bicinchoninic acid)법검측bcl-2기인표체단백농도，수거진행통계학분석。결과 A조세포조망솔교D조저(u=2.16，0.02<P<0.05), B조교E조저（u=2.38，0.01<P<0.02），C 조교B조저(u=2.27，0.02<P<0.05)급교F조저(u=2.83，0.02<P<0.05),통계학분석유현저성차이。A조bcl-2기인표체단백농도고우D조（t=2.87，0.005<P<0.002）， B조고우E조（t=2.75，0.01<P<0.05），C조고우B조（t=2.16，0.02<P<0.05）급F조（t=2.23，0.02<P<0.05）,통계학분석유현저성차이。결론 bcl-2기인급NGF균대정상신경세포적조망유억제작용，능구증강신경세포적항손상능력，이자연합응용시구유협동작용。
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 MOI.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.Group D: PC12 cells with slow virus plasmid. Group E: PC12 cells with slow virus plasmid, which were damaged by H2O2. Group F: PC12 cells with slow virus plasmid, which were damaged by H2O2., were treated with NGF.The rate of apoptosis was detected by flow cytometry .By BCA(bicinchoninic acid) method,the proteinum concentration of bcl-2 gene expression was detected.Results The apoptosis rate of group A was lower than that of groupD.The apoptosis rate of group B was lower than that of groupE.The apoptosis rate of group C was lower than that of groupB and was lower than that of groupF.Protein concentrations of bcl-2 gene expression of group A was higher than that of group D.Protein concentrations of bcl-2 gene expression of group B was higher than that of group E Protein concentrations of bcl-2 gene expression of group C was higher than that of group B and was higher than that of group F.There were statistical y significant difference between two groups (p<0.05).Conclusion Both bcl-2 gene and NGF can inhibit apoptosis of normal nerve cells and can enhance resistance ability of nerve cellto be damaged. There were synergia abaissement role on the apoptosis of nerve cells when they were used together.