[目的]明确GABPα在鱼藤酮诱导的PC12细胞氧化应激及凋亡中的作用。[方法]利用鱼藤酮制备PC12细胞氧化应激模型，通过脂质体转染的方法在PC12细胞中过表达GABPα。利用Western Blot检测GABPα的表达，通过可见分光光度法和微量法分析氧化应激标记物NO和MDA水平，抗氧化标记物GSH的水平和SOD的活性，利用流式细胞仪检测细胞凋亡。[结果]与对照组相比，鱼藤酮处理后，PC12细胞的活力明显降低(46。71％±1。7％vs 99。88％±0。649％)，NO 和 MDA 的水平明显增高(0。285±0。004 vs 0。151±0。003，0。115±0。003 vs 0。044±0。002)，GSH 的水平及 SOD 的活性显著下降(11。53±0。572 vs 22。86±1。338，0。161±0。008 vs 0。315±0。026)，细胞的凋亡明显增加(30。26％±2。359％vs 3。037％±0。043％)。在PC12细胞过表达GABPα并用鱼藤酮处理后，与空载体组相比，PC12细胞的活力明细增高(62。21％±2。344％vs 47。65％±3。228％)，氧化应激产物NO和MDA的水平明显下降(0。194±0。004 vs 0。268±0。003，0。075±0。005 vs0。112±0。004)，GSH 的水平及 SOD 的活性明显升高(18。03±1。18 vs 11。22±0。474，0。242±0。006 vs 0。159±0。003)，细胞的凋亡显著降低(7。047％±0。788％vs 31。98％±2。929％)。[结论]GABPα在PC12细胞中可通过抑制鱼藤酮诱导的氧化应激和细胞凋亡而起到神经保护作用。
GABPα inhibits the oxidative stress and apoptosis induced by rotenone in PC12 cells
[Objective]To determine the role of GABPα in rotenone-induced oxidative stress and apoptosis in PC 12 cells.[Method]The oxidative stress model of PC12 cell was built by rotenone treatment.The GABPα gene was overexpressed in PC12 cells by liposome transfection.The expression of GABPα was detected by Western Blot.The level of oxidative stress marker(NO and MDA)and antioxidant marker(GSH and SOD)were analyzed in PC12 cells by visible spectrophotometry and micro-determination.The PC12 cell apoptosis was detected using flow cytometry.[Result]Compared with the control group,the via-bility of PC 12 cells was significantly reduced(46.71％±1.7％vs 99.88％±0.649％).Meanwhile,the levels of NO(0.285±0.004 vs 0.151±0.003)and MDA(0.115±0.003 vs 0.044±0.002)as well as the cell apoptosis(30.26％±2.359％vs 3.037％±0.043％)were obviously increased after the treatment with rotenone.However,the level of GSH(11.53±0.572 vs 22.86±1.338)and the activity of SOD(0.161±0.008 vs 0.315±0.026)were decreased.After the GABPα gene was ex-pressed in PC 12 cells,the viability of PC12 cells(62.21％±2.344％vs 47.65％±3.228％),the level of GSH(18.03±1.18 vs 11.22±0.474)and the activity of SOD(0.242±0.006 vs 0.159±0.003)were increased compared with the empty vector group in the presence of rotenone.However,the levels of NO(0.194±0.004 vs 0.268±0.003)and MDA(0.075±0.005 vs 0.112±0.004)as well as the cell apoptosis(7.047％±0.788％vs 31.98％±2.929％)were decreased.[Conclusion]GABPα can exert neuroprotective effects in PC 12 cells by inhibiting the rotenone-induced oxidative stress and cell apoptosis.
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