转染PHGPx基因对Aβ及H_2O_2诱导PC_(12)细胞凋亡的保护作用

Transfection of PHGPx gene can protect PC_(12) cells from apoptosis induced by amyloid beta-peptide or hydrogen peroxide

目的 观察转染磷脂过氧化氢谷胱甘肽过氧化物酶 (PHGPx)基因对神经毒物质Aβ及H2 O2 诱导细胞凋亡的保护作用。方法 通过基因转染和筛选得到稳定高表达PHGPx基因的PC1 2 细胞 ,加入神经毒物质Aβ及H2 O2 ,采用电镜、DNA片段化测定、MTT比色微量分析及流式细胞凋亡率分析等方法观察转基因后抗逆变致凋亡的能力。结果 加入神经毒物质Aβ或H2 O2 后 ,未转染PHGPx基因的PC1 2 细胞出现凋亡的特征性形态学改变 ,电泳可见断裂的DNA片段 ,流式细胞仪检测细胞凋亡率较转染组明显增加。MTT比色微量分析显示细胞活力下降 (均P <0 0 1 )。结论 Aβ及H2 O2 能够诱导神经细胞凋亡。转染PHGPx基因能对抗Aβ、H2 O2 的神经毒性 ,保护Aβ、H2 O2 所致的神经细胞损伤凋亡 。

Objective To observe the protective effects of phospholipid hydroperoxide glutatione peroxidase (PHGPx) gene transfection against PC 12 cells apoptosis induced by amyloid beta-peptide or hydrogen peroxide. Methods We transfected and selected the PC 12 cells which can stably overexpress PHGPx gene, then exposed the cells to Aβ and hydrogen peroxide, measured the changes of cells apoptosis by electric microscope, MTT assay, DNA electrophoretic analysis and flow cytometric DNA analysis.Results Exposure of PC 12 cells without PHGPx gene to Aβ or hydrogen peroxide resulted in characteristic changes of apoptosis, including morphologic changes, DNA fragmentation and an increase in the apoptosis rate. While in the cells transfected with PHGPx genes, there was no such remarkable changes when treated with Aβ or hydrogen peroxide.Conclusions Aβ and hydrogen peroxide could induce neural cells apoptosis. Transfection of PHGPx genes can protect cells from apoptosis induced by Aβ or hydrogen peroxide. This method may be a new gene therapeutic method to treat neurodegenerative diseases such as AD.