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目的 研究基因bcl 2对神经元的生物活性作用。 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和原位免疫组织化学检测外源基因表达 ;10 μmol L、5 0 μmol L和10 0 μmol 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 μmol L、5 0 μmol L和 10 0 μmol L顺铂处理后 ,实验A组存活的细胞数分别为 2 76± 13、185± 11和 10 8± 10 ,而对照B组中存活的细胞数分别为 10 0± 9、12± 3和 2± 2 ,两者相比有显著性差异 ;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 2 5 79%明显减少。 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
Objective To study the biological activity of gene bcl 2 on neurons. Methods The eukaryotic expression vector pcDNA3 bcl 2 was constructed. The recombinant plasmid was introduced into PC12 cells by lipofectamine. The expression of foreign gene was detected by Western blot and in situ immunohistochemistry. The expression of exogenous gene was detected by 10 μmol L, 50 μmol L and 100 μmol L Platinum treated group A cells transfected with recombinant plasmids. Cells in control group B transfected with empty plasmid were also treated under the same conditions. After 72 hours, the number of surviving cells in both groups was compared. Cell cycle index was analyzed by flow cytometry. Results The eukaryotic expression vector pcDNA3 bcl 2 was successfully constructed and the cell clone stably expressing Bcl 2 was obtained by liposome-mediated method. After treated with 10 μmol L, 50 μmol L and 100 μmol L cisplatin, The number of surviving cells was 2 76 ± 13,185 ± 11 and 10 8 ± 10, respectively, while the number of surviving cells in control group B was 10 ± 9,12 ± 3 and 2 ± 2, respectively Significant difference; flow cytometry showed that the percentage of S phase cells in experimental group A was 81 1%, which was significantly lower than that of control group B 25 79%. Conclusions bcl 2 can antagonize the damaging effect of cisplatin on neurons and promote the survival of nerve cells, and its neuroprotective mechanism may be accomplished by regulating the cell cycle