神经妥乐平对H2O2致PC12细胞损伤的保护作用  

Neuroprotective effect of Neurotropin on H2O2-induced injury in pheochromocytoma cells

作　　者：方文丽[1] 范胜诺[1] 廖旺[1] 郑雨秋 廖劭伟 刘军[1] 

机构地区：[1]中山大学孙逸仙纪念医院神经科,广州510120

出　　处：《国际医药卫生导报》2017年第6期771-785,共15页International Medicine and Health Guidance News

基　　金：2013年国家自然科学基金（81372919）;广东省自然科学基金项目（2014A030313016）

摘　　要：目的 探索神经妥乐平对H2O2诱导的PC12细胞氧化应激损伤的影响及其潜在机制。方法 用CCK-8法检测细胞存活率，以流式细胞术检测细胞氧化损伤的发生、细胞内活性氧的生成及线粒体膜电位的变化，荧光显微镜观察细胞内活性氧的产生，qRT-PCR 测定 Caspase-3、Bax 和 Bcl-2 mRNA 的表达。结果 PC12 细胞存活率随 H2O2 浓度的增加而逐渐下降。其中，450μM H2O2 处理细胞 24 h 后细 胞存活率、凋亡率、坏死率明显降低；细胞内活性氧水平表达明显升高；线粒体膜电位JC-1红/绿荧光比值下降；Bax 和 Caspase-3 的 mRNA 表达升高，而 Bcl-2 的 mRNA 表达下降，以上指标与对照组相比， 差异有统计学意义 （P ＜ 0.05）。而预先给予 0.01 UN/ml 的 NTP 处理细胞 12 h 可明显提高细胞存活率，降 低细胞凋亡率和坏死率，减少细胞内活性氧生成并提高线粒体膜电位，抑制Bax和Caspase-3的mRNA表 达，促进 Bcl-2 mRNA 的表达，以上指标与 H2O2 组相比，差异有统计学意义 （P ＜ 0.05）。结论 NTP 能抑 制H2O2诱导的PC12细胞损伤，其神经细胞保护作用可能与其降低细胞内活性氧水平、维持线粒体膜电位的高能状态和抑制促凋亡基因表达、促进抗凋亡基因表达有关。Objective To investigate the effect of Neurotropin on H202-induced oxidative stress injury in pheochromocytoma （PC 12） cells and its potential mechanism. Methods The CCK-8 assay was applied to measure the cell viability while the flow cytometry was used to detect the occurrence of cells oxidative injury, the level of intracellular reactive oxygen species （ROS） and changes of mitochondrial membrane potential. The ROS was observed by fuorescence microscope, and the mRNA expressions of Caspase-3, Bax, and Bcl-2 were detected by immunofluorescence and quantitative real time polymerase chain reaction （qRT-PCR）. Results As the increase of H2O2 concentration, the survival rate of PC12 cells was decreased gradually. The treatment with 450 pM of H202 for 24 h induced the cell viability, apoptosis and necrosis significantly and increased the level of the ROS evidently; the red/green fluorescence ratios of JC-1 were decreased in the mitochondrial membrane potential; the mRNA expressions of Bax and Caspase-3 were significantly increased, while the mRNA expression of Bcl-2 was decreased （P〈0.05, compared with the control group）. In contrary, with pretreatment of Neurotropin （0.01 UN/ml） for 12 h, it obviously enhanced the cell viability and mRNA expressions of Bcl-2, decreased the rates of apoptosis and necrosis, lessened the accumulation of the ROS, increased the mitochondrial membrane potential, and inhibited the expressions of Bax and Caspase-3 （P〈0.05, compared with the H2O2 group）. Conclusion Neurotropin can inhibit the H2O2-induced injury in PC12 cells. Its neuroprotective role may relate to decreasing the level of the ROS, maintaining the high energy of the mitochondrial membrane potential, inhibiting the expression of pro- apoptotic gene expression and promoting anti-apoptotic gene expression.

关 键 词：神经妥乐平 PCI2细胞 氧化应激损伤 线粒体膜电位 凋亡 

分 类 号：R96[医药卫生—药理学]