基因bcl-2对神经元生物活性的保护作用(英文)

The protective effect to neuronal bioactivity by bcl-2gene

目的研究基因bcl-2对神经元的生物活性作用,为神经系统疾病的保护治疗提供有效的途径。方法构建真核表达载体pcDNA3-bcl-2,采用脂质体介导将重组质粒导入PC12细胞,WesternBlot检测外源基因表达;10,50和100μmol/L顺铂处理转染重组质粒的实验A组细胞,同样条件处理转染空质粒的对照B组细胞,72h后比较两者存活的细胞数;流式细胞仪检测分析两者的细胞周期指数。结果成功构建了真核表达载体pcDNA3-bcl-2,并用脂质体介导的方法获得了稳定表达Bcl-2的细胞克隆;10,50和100μmol/L顺铂处理后,实验A组存活的细胞数分别为276±13,185±11和108±10,而对照B组中存活的细胞数分别为100±9,12±3和2±2,两者相比差异有显著性意义;流式细胞仪检测显示,实验A组S期细胞所占百分比为8.81%比对照B组25.79%明显减少(χ2=22.53,P<0.01)。结论bcl-2能够拮抗顺铂对神经元的损害作用,促进神经细胞存活,其神经保护作用机制可能通过调控细胞周期来完成。

AIM:To investigate the biological effects of bcl-2gene o n neurons and to provide effective methods for treat ment for nervous system dis eases.METHODS:The recombinant expression plasmid pcDNA3-bcl-2was construct ed from pSFFV-bcl-2,then it was introduced into PC12cell line by liposome method.Western blottin g were applied to detect the exogenou s gene expression .The two groups of cells(group A:PC12transfected by pcDNA3-bcl-2and gro up B:PC12transfected by pcDNA3)were exposed to Cisplatin with the concentra tion of 10,50and 100μmol /L.72hours later,the survival cells in tw o grou ps were examined.Cell cycle indexes between these two groups were also s tudied by FCM.RESULTS:The recombinant expression plasmid pcDNA3-bcl-2was c on-structed successfully and PC12cell line transfected by the plasmid coul d express Bcl-2protein effectively.After exposed to 10,50and 100μmol /LCisplatin,the surviving cells in group A were 276±13,185±11and 108±10respectively,which increased much more than those in group B while they were 100±9,12±3and 2±2accordingly and significant differences were found.Compared with the control (25.79%),there was a significant de-crease of cells from the S phase in PC12with bcl-2gene(8.81%,χ 2 =22.53,P<0.01).