miR-204对癫神经元中BDNF/TrkB信号通路的影响  

The Study of miR-204 Regulates BDNF/TrkB Expression in Epileptic Neurons

作　　者：常伟[1] 潘立平[2] 吴秋静 宋毅军[2] 

机构地区：[1]天津医科大学, 300070 [2]天津医科大学总医院神经内科, 300070

出　　处：《天津医药》2014年第3期214-216,I0005,共4页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:91132722、81071044)

摘　　要：目的研究海马神经元癫模型中转染miR-204后对脑源性神经营养因子(BDNF)与TrkB通路调控作用的影响。方法取原代培养7d后的细胞,分为正常组、正常+BDNF组、癫组、癫+BDNF组、正常转染miR-204组、癫转染miR-204组、癫转染miR-204+BDNF组。癫组用无镁液处理3 h制作癫模型。制备成功miR-204慢病毒表达载体。采用免疫荧光、膜片钳及免疫印迹技术鉴定及观察miR-204对BDNF/TrkB通路表达的影响。结果 TrkB蛋白的磷酸化水平:正常+BDNF组高于正常组;癫+BDNF组低于正常+BDNF组,高于癫组;癫+miR-204+BDNF组高于癫+BDNF组和癫+miR-204组。结论 BDNF及miR-204可以改善BDNF/TrkB受抑制的状态,从而在癫疾病的缓解中可能发挥重要作用。Objective To study the effect of miR-204 on BDNF/TrkB signaling and pathogenesis on the neuron model of epilepsy. Methods Primary hippocampal neurons were cultured in vitro for 7 days, and were divided into control group, control＋BDNF group, epilepsy group, epilepsy＋BDNF group , control＋miR204 group, epilepsy＋miR204 group and ep-ilepsy＋miR204＋BDNF group. The epilepsy model of hippocampal neurons was established by being exposed to Mg2＋free me-dia for 3 hours. The miR-204 lentivirus vector was constructed. The effect of miR-204 on BDNF/TrkB expression was detect-ed by immunohistochemistry, patch clamp and Western blot technique. Results Compared with the control group, the TrkB phosphorylation level was higher in control＋BDNF group. The TrkB phosphorylation level was lower in epilepsy＋BDNF group than that of control＋BDNF group, but it was higher than that of epilepsy group. The TrkB phosphorylation level was higher in epilepsy＋miR204＋BDNF group than that of epilepsy＋BDNF group and epilepsy＋ miR204 group. Conclusion BDNF and miR-204 can improve the inhibitory condition of BDNF/TrkB signaling and may play an important role in alleviat-ing epilepsy disease.

关 键 词：癫痫 海马 神经元 脑源性神经营养因子 miR-204 miR-204 

分 类 号：R749.1[医药卫生—神经病学与精神病学]