氧化应激后大鼠神经元内Nogo-A的表达变化

Investigation of expressional change and function of Nogo-A in neurons after oxidative stress

目的 :探讨大鼠大脑皮层神经元受到氧化应激后Nogo A表达的变化及其可能的作用 .方法 :体外原代培养大鼠大脑皮层神经元 ,给予不同浓度H2 O2 处理 8h后 ,倒置相差显微镜下观察神经元的形态学变化、Hoechst33342和PI染色观察凋亡和坏死情况 ,然后Westernblot观察Nogo A的表达变化 .结果 :大鼠皮层神经元受到 10和 10 0 μmol/LH2 O2刺激后 ,神经元出现胞体回缩、变圆甚至崩解 ,突起断裂、脱落等细胞受损的现象 ;Hoechst33342和PI染色观察到 1μmol/LH2 O2 时神经元凋亡和坏死比例与对照组相比没有差别 ,而 10和 10 0 μmol/LH2 O2 刺激后 ,神经元凋亡和坏死比例明显增加 (P <0 .0 5 ) .Westernblot结果显示 ,1μmol/LH2 O2 时No go A的表达量与对照组相比无明显差异 ,而在 10和 10 0μmol/L时Nogo A的表达量明显升高 (P <0 .0 5 ) .结论 :氧化应激可以上调神经元Nogo A的表达 .

AIM: To investigate the change of Nogo-A expression in cerebral cortex neurons after oxidative stress and its possible functions. METHODS: Primarily cultured cerebral cortex neurons in rats were treated with H 2O 2 for 8 h and the morphology of neurons was observed. Hoechst33342 and PI staining were performed to detect apoptosis and necrosis of neurons, and Nogo-A expression was detected by Western blot. RESULTS: We observed that soma of neurons condensed, rounded, even collapsed, and processes disrupted and shed after treatment with 10 μmol/L and 100 μmol/L H 2O 2. Hoechst33342 and PI staining showed that, compared with that of control group, the percentage of apoptosis and necrosis did not increase after neurons were treated with 1 μmol/L H 2O 2, while it significantly increased after neurons were treated with 10 μmol/L and 100 μmol/L H 2O 2 (P<0.05). Western blot analysis showed that Nogo-A expression had no difference with that of control group after neurons were treated with 1 μmol/L H 2O 2, whereas was significantly higher than control group after neurons were treated with 10 μmol/L and 100 μmol/L H 2O 2 (P<0.05). CONCLUSION: Oxidative stress may up-regulate the expression of Nogo-A in neurons, and Nogo-A is presumed to be involved in the pathological processes of CNS degenerative diseases.