Notch信号通路在新生鼠高氧暴露致少突胶质细胞成熟障碍中的作用.doc

10.16016/j.1000-5404.201412058 Notch信号通路在新生鼠高氧暴露致少突胶质细胞成熟障碍中的作用 刘光建，郭 冉，杜 敏，刘 杨，刘剑霞，徐 颖 (400014重庆，重庆医科大学附属儿童医院麻醉科，儿童发育疾病研究省部共建教育部重点实验室，儿科学重庆市重点实验室) [摘要] 目的 探讨Notch信号通路在新生鼠高氧暴露致少突胶质细胞成熟障碍中的作用。 方法 取C57BL/10J新生3 d（P3）小鼠64只，分为空气对照组（C）、空气+DAPT组（C＋DAPT）、高氧组（H）、DAPT预处理高氧暴露组（H+DAPT）；H组予以高氧暴露48h，H+DAPT组采用Notch信号特异抑制剂DAPT（10mg/kg）腹腔注射1 h后高氧暴露48 h。生后5 d（P5）断头取脑。Real-time PCR检测Notch1、Jagged1、Hes1及Hes5表达变化。生后12 d（P12）取脑，HE染色观察脑白质形态学变化；免疫荧光化学染色检测成熟少突胶质细胞（OL）表达；Western blot定量检测MBP蛋白的表达量。 结果 与C组比较，H组小鼠脑组织明显水肿，Notch1、Jagged1、Hes1及Hes5 mRNA的表达均增高（P0.05）；与H组比较，H+DAPT组Notch1、Jagged1mRNA表达明显增强（P0.05），Hes1及Hes5 mRNA 表达显著降低（P0.05）；免疫荧光和Western blot结果均显示H组MBP表达低于C组（P0.05）；而与H组比较，H+DAPT组MBP表达显著增多（P0.05）。结论 Notch信号通路参与了高氧暴露致新生鼠脑白质损伤过程，其机制可能与影响少突胶质细胞的成熟有关。 [关键词] 高氧；DAPT；脑损伤；Notch信号通路；少突胶质细胞成熟 [基金项目] 国家临床重点专科建设项目(2013544) [通信作者]徐颖，E-mail:xuyingxy3066@126.com Effects of Notch signaling pathway on hyperoxia-induced oligodendrocyte maturation disorders in neonatal mouse Liu Guangjian,Guo Ran,Du Ming,Liu Yang,Liu Jianxia, XuYing(Department of Anesthesiology; Key Laboratory of Developmental Diseases in Childhood Chongqing and Ministry of Education，Childrens Hospital，Chongqing，400014，China) [Abstract] Objective To investigate the effects of Notch signaling pathway on hyperoxia-induced oligodendrocytes maturation disorders in neonatal mouse brain tissue. Methods Sixty-four C57BL/10J neonatal mouses are randomly divided into control group, control+DAPT (10mg/kg) group, hyperoxia group (hyperoxia exposure 48h), hyperoxia+DAPT group (hyperoxia exposure 48h after 10mg/kg DAPT treatment). All neonatal mouses were killed and brain was removed to the following observation and detection: at P5, the mRNA expression variation of Notch1, Jagged1，Hes1 and Hes5 by Real-time PCR methods；at P12, oligodendrocytes (OL) were identified by immunofluorescence staining; Myelin basic protein (MBP) protein expression was detected by western blot assay. Results Hyperoxia group showed brain injury characterized by inh