丁苯酞对肌萎缩侧索硬化模型小鼠的生存期脊髓前角运动神经元Nrf2及HO-1表达

目的探讨丁苯酞(NBP)对SOD1~(G93A)转基因小鼠的生存期及脊髓前角运动神经元Nrf2、HO-1表达的影响及机制。方法肌萎缩侧索硬化(ALS)模型SOD1~(G93A)转基因小鼠24只,随机分为NBP组(n=12雌雄各半)和安慰剂组(n=12雌雄各半)选同窝SOD1~(G93A)阴性小鼠作为阴性对照(n=12雌雄各半)。于6w龄起分别给予NBP组NBP 180mg·kg~(-1)·d~(-1),安慰剂组玉米油10mL·kg~(-1)·d~(-1)灌胃,至终末期取材。观察两组的发病时间及生存期。通过甲苯胺蓝染色比较终末期两组小鼠脊髓前角运动神经元数目,通过免疫组织化学染色、蛋白印迹法观察干预前后脊髓前角Nrf2、HO-1的分布、表达情况。结果相比安慰剂组,NBP组可推迟转基因小鼠的发病时间(P0.05)显著延长小鼠生存期(P0.01);终末期残存的脊髓前角运动神经元数量明显增加(P0.01)。免疫组织化学染色结果显示,与安慰剂组相比,NBP组Nrf2在胞质内具有较强的免疫反应性差异有统计学意义(P=0.004)。NBP组HO-1在胞质内具有较强的免疫反应性差异无统计学意义(P=0.275)。蛋白印迹结果显示,相比安慰剂组,NBP组Nrf、HO-1蛋白的表达均明显增加(P均0.05)。结论NBP可以推迟SOD1~(G93A)转基因小鼠的发病时间并显著延长生存期,增加细胞质内Nrf2、HO-1的表达。

Effects of dl-3n-butylpthalide on lifespan and Nrf2 and HO-1 expression in the mice model of amyotrophic lateral sclerosis

Objective To study the effects of dl-3n-butylpthalide(NBP) on the lifespan in the SOD1G 93A transgenic mice and NFE2-related factor 2 ( Nrf2) and Heme oxygenase-1 ( HO-1) expression in the spinal cord anterior horn.Methods Twenty-four SOD1G 93A transgenic mice for amyotrophic lateral sclerosis model were randomly divided into NBP group and placebo group (n=12,6 males and 6 females).Another 12 SOD1G 93A negative littermate mice were used as negative control group .NBP 180mg? kg-1 ? d-1 and corn oil 10mL? kg-1 ? d-1 were administered through oral gavages respectively at 6 weeks.Onset and lifespan were observed in the two groups .The motor neuron counts in the anterior horn of spinal cords were studied in terminal stage of both groups by toluidine blue staining,the expression and distribution of Nrf2 and HO-1 in the anterior horn of spinal cords were investigated by immunohistochemical ABC and Western blotting methods .Results Compared with the placebo group ,the NBP group could delay the onset(P<0.05) and significantly extend the lifespan of the transgenic mice (P<0.01).The NBP group reserved obviously more neurons in terminal stage (P<0.01).The immunohistochemistry results showed that NBP could increase Nrf2 expression in neuron cytoplasm (P=0.004),and NBP group had more HO-1 positive neurons by immunohistochemistry method compare with placebo group (P =0.275 ).Furthermore,Nrf2 and HO-1 protein in NBP group revealed significantly increase by Western blotting (P<0.05).Conclusion NBP can delay the onset time and extend the lifespan of SOD1G93A transgenic mice.Besides,it can reduce the loss of motor neurons in the anterior horn of spinal cord and increase expression of Nrf 2/HO-1 in neuron cytoplasm.