丙戊酸通过CREB上调脑红蛋白并保护N2a细胞免受双氧水诱导的神经损伤  

Valproic acid induces neuroglobin protein by CREB and protects N2a cells against H_2O_2-induced neurotoxicity

作　　者：刘宁[1,2,3] 寻禹 李亚丹 王婷婷[2,3] 钟爱军 姚亮元 袁秀菊 向双林[2,3] 

机构地区：[1]湖南千金湘江药业,湖南株洲412000 [2]湖南师范大学生命科学学院,湖南长沙410000 [3]湖南省生物发育工程与新产品研发协同创新中心,湖南长沙410000

出　　处：《中国药理学通报》2014年第5期619-623,共5页Chinese Pharmacological Bulletin

基　　金：国家重点基础研究发展计划(973计划)资助项目(No 2010CB529900);湖南省教育厅重点项目(No 11A072);湖南省属科研机构技术创新发展专项(No 2010TF2014);长沙市科技局重点项目(No K1205221-31)

摘　　要：目的研究丙戊酸对脑红蛋白表达的影响和机制,及其对双氧水诱导的神经损伤的保护作用。方法 Western blot、RT-PCR和荧光素酶活性实验检测丙戊酸对小鼠和人脑红蛋白表达的影响;荧光素酶活性实验分析CREB在丙戊酸诱导脑红蛋白启动子活性过程中的作用;MTT实验分析丙戊酸对双氧水诱导的神经损伤的保护能力。结果发现丙戊酸上调小鼠和人脑红蛋白的蛋白和mRNA水平,并能上调脑红蛋白基因的启动子活性;CREB特异性抑制剂KG-501和CREB显性负突变体KCREB均抑制丙戊酸诱导脑红蛋白基因的启动子活性;丙戊酸可保护N2a神经瘤母细胞免受双氧水诱导的损伤。结论 CREB介导了丙戊酸诱导的脑红蛋白表达上调;脑红蛋白可能在丙戊酸保护N2a细胞免受氧化应激诱导的神经损伤过程中扮演重要作用。Aim To investigate the effect and mechanism of valproic acid on neuroglobin expression, and the neuroprotective role of valproic acid against H2O2- induced neurotoxicity. Methods Western blot, RT- PCR and luciferase assay were used to detect the protein levels, mRNA levels and promoter activity of mouse and human neuroglobin induced by valproic acid. Luciferase assay was used to investigate the role of transcription factor CREB in the up-regulation of neuroglobin by valproic acid. MTT assay was used to evaluate the effect of valproic acid against H2O2-induced neurotoxicity. Results VPA treatment markedly increased the protein levels, mRNA levels and promoter activity of Ngb in SKNSH cells. CREB mouse N2a cells and human specific inhibitor KG501 or CREB dominant negative mutant KCREB attenuated VPA-induced Ngb promoter activity. VPA could protect N2a ceils from H2O2-induced neurotoxicity. Conclusion CREB mediates VPA-induced Ngb up-regulation, which may contribute to the neuroprotective effects of VPA in oxidative stress in neurons.

关 键 词：丙戊酸 脑红蛋白 CREB N2a SKNSH 双氧水 神经损伤 

分 类 号：R-332[医药卫生] R322.81