Notch 2在肌萎缩侧索硬化症SOD1^(G93A)转基因小鼠脊髓内的表达变化  

Dynamic expression of Notch 2 in the spinal cords of amyotrophic lateral sclerosis SOD1^(G93A) transgenic mice

作　　者：张皓云[1] 薄其付[2] 杜红梅[3] 陈燕春[3] 郑昕蕊 于丽[3] 管英俊[3] 

机构地区：[1]潍坊医学院基础医学概论教研室,潍坊261053 [2]潍坊医学院附属医院肿瘤科,潍坊261053 [3]潍坊医学院组织胚胎学教研室,潍坊261053

出　　处：《中国组织化学与细胞化学杂志》2017年第5期439-444,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金(81271413);山东省自然科学基金项目(ZR2014JL016;ZR2015HL047);山东省高等学校科技计划项目(J15LK10)

摘　　要：目的明确肌萎缩侧索硬化症(amyotrophic lateral sclerosis,ALS)SOD1G93A转基因小鼠脊髓内Notch 2表达水平与ALS病程的关系。方法应用Notch基因阵列表达谱检测Notch信号通路相关分子在表达人突变SOD1基因的SOD1G93A转基因ALS小鼠及野生型鼠脊髓内的表达差异。应用real-time PCR、免疫组织化学染色、Western blot方法检测Notch 2在不同阶段各组小鼠脊髓内的表达变化。结果 Notch基因阵列表达谱聚类分析结果显示Notch 1、Notch 2在95d、122d SOD1G93A鼠脊髓内均显著增高,而Notch 4仅在122d SOD1G93A鼠升高明显。Notch下游靶基因HES1在108d SOD1G93A鼠脊髓内显著降低。RT-PCR结果证实Notch 2在SOD1G93A鼠脊髓内的表达变化。免疫组织化学标记显示Notch 2在SOD1G93A鼠、野生型鼠脊髓各部均有表达,Western blot结果显示Notch 2蛋白表达在95d SOD1G93A鼠增高显著。结论 Notch 2信号分子的表达随ALS疾病进展,在脊髓退变过程中变化显著,可能参与ALS疾病进程。Objective To investigate the relationship between Notch2 expression and disease stage in the spinal cords of amyotrophic lateral sclerosis （ALS） SOD1G93A transgenic mice. Method Using Notch signaling pathway polymerase chain reaction array to determine the expression levels of Notch signaling molecules in SOD1G93A transgenic mice and wild type mice. The Notch2 expression in the spinal cord of mice at different stages was further evaluated by real-time PCR, immunohistochemical staining and Western blot. Results Notch Signaling Pathway PCR Array showed that compared to wild-type mice, Notch 1 and Notch2 were significantly up-regulated in the spinal cords of SOD1G93A mice at 95 d and 122 d, while increased Notch4 expression was observed only at 122 d. The downstream target gene Hes1 was significantly reduced at 108 d. RT-PCR confirmed the increased Notch2 expression at mRNA level. Immunohistochemical staining indicated that Notch2 was expressed throughout the spinal cord of both SOD1G93A and wild type mice. Western blot results showed a significant up–regulation of Notch2 in SOD1G93A mice compared to wild type mice at 99 d. Conclusion Our findings provide evidence that the expression of Notch2 signaling molecules was strongly associated with ALS disease progression and spinal cord neuronal degeneration in adult SOD1G93A mice, which suggested it may be involved in ALS pathogenesis.

关 键 词：肌萎缩侧索硬化症 转基因小鼠 NOTCH信号通路 NOTCH2 脊髓 

分 类 号：R742[医药卫生—神经病学与精神病学]