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目的:检测不同疾病阶段肌萎缩侧索硬化症(ALS)转基因小鼠脊髓内Nrf2及HO-1表达变化,以探究抗氧化应激能力变化与ALS发病的关系。方法:应用免疫荧光双重标记和Western Blot技术检测Nrf2及其下游调节蛋白HO-1在ALS发病早期(95 d)、发病晚期(122 d)脊髓中的表达差异。结果:与同窝别野生型鼠相比较,免疫荧光双重标记结果显示:95 d、122 d ALS鼠脊髓腰段内Nrf2~+/GFAP~+星形胶质细胞增多;虽与同时间点同窝别野生型鼠比较,ALS转基因鼠脊髓中Nrf2蛋白表达略有升高,但差异无统计学意义。HO-1蛋白表达在ALS早期95 d趋势与Nrf2相同,前角神经元内表达较多;至122 d转基因鼠脊髓内HO-1~+表达多集中于星形胶质细胞内,且脊髓内HO-1蛋白总量较同窝别野生型鼠显著升高(P<0.05)。结论:ALS运动神经元退变可能与星形胶质细胞激活后Nrf2及HO-1参与的氧化应激机制有关。
Objective: To detect the expression of Nrf2 and HO-1 in the spinal cord of patients with Amyotrophic Lateral Sclerosis (ALS) transgenic mice in different stages of disease to explore the relationship between the changes of antioxidant stress and the pathogenesis of ALS. Methods: The expression of Nrf2 and its downstream regulatory protein HO-1 in the early stage of ALS (95 d) and late onset (122 d) spinal cord were detected by immunofluorescence double labeling and Western Blot. Results: Compared with the wild type littermates, immunofluorescence double labeling results showed that the number of Nrf2 ~ + / GFAP ~ + astrocytes in spinal cord increased in 95 d and 122 d ALS mice, Compared with wild type mice, the expression of Nrf2 protein in the spinal cord of ALS transgenic mice slightly increased, but the difference was not statistically significant. The expression of HO-1 protein in the early stage of ALS was similar to that of Nrf2 at 95 days, and was more in the anterior horn neurons. The expression of HO-1 ~ + in the spinal cord of 122 days transgenic mice was mainly localized in astrocytes, The total amount of HO-1 protein was significantly higher than that of wild-type mice (P <0.05). Conclusion: The degeneration of ALS motor neurons may be related to the oxidative stress mechanisms involved in the activation of astrocytes after Nrf2 and HO-1.