TrkB不同亚型对癫海马神经元BDNF/TrkB信号通路调控的研究  

An Experimental Study of the Regulation of BDNF/TrkB Signal Pathway by Different Isoforms of TrkB in Epileptic Hippocampal Neurons

作　　者：吴秋静 常伟[1] 潘立平[1] 宋毅军[1] 赵文[1] 

机构地区：[1]天津医科大学总医院神经内科,300052

出　　处：《天津医药》2014年第5期406-409,共4页Tianjin Medical Journal

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

摘　　要：目的探讨癫海马神经元中酪氨酸激酶受体B(TrkB)不同亚型对脑源性神经营养因子(BDNF)/TrkB信号通路的调控机制。方法取原代培养7 d后的海马神经元,分为钙调蛋白抑制剂(ALLN)组和翻译抑制剂(An-isomycin)两大组。ALLN组又分为正常组、正常+BNDF组、癫组、癫+BDNF组、正常+ALLN组、癫+ALLN组和癫+ALLN+BDNF组;Anisomycin组又分为正常组、正常+BDNF组、癫组、癫+BDNF组、正常+Anisomycin组、癫+Anisomycin组和癫+Anisomycin+BDNF组。免疫荧光鉴定海马神经元,无镁液处理制备癫模型,电生理鉴定细胞痫样放电,免疫印迹技术检测各组TrkB和磷酸化TrkB(p-TrkB)蛋白的表达变化。结果 (1)ALLN组:正常+BDNF组p-TrkB/TrkB灰度值高于正常组;癫+BDNF组高于癫组,低于正常+BDNF组;癫+ALLN+BDNF组低于癫+BDNF组,与癫+ALLN组差异无统计学意义。(2)Anisomycin组:正常+BDNF组p-TrkB/TrkB灰度值高于正常组;癫+BDNF组高于癫组,低于正常+BDNF组;癫+Anisomycin+BDNF组高于癫+BDNF组和癫+Anisomy-cin组。结论通过Anisomycin降低TrkB.T表达可改善癫状态下BDNF/TrkB受抑制的状态,通过ALLN升高TrkB.FL表达无法改善其抑制状态。Objective To investigate the mechanism of brain derived neurotrophic factor （BDNF） regulated by differ-ent isoforms of tyrosine kinase receptor B （TrkB） in epileptic hippocampal neurons. Methods Primary hippocampal neu-rons were cultured in vitro for 7 days, and divided into two groups, ALLN （calcineurin inhibitor） group and Anisomycin （trans-lation inhibitor） group. ALLN group included control group, control＋BDNF group, epilepsy group, epilepsy＋BDNF group, control＋ALLN group, epilepsy＋ALLN group and epilepsy＋ALLN＋BDNF group. Anisomycin group was sub-divided into con-trol group, control＋BDNF group, epilepsy group, epilepsy＋BDNF group, control＋Anisomycin group, epilepsy＋Anisomycin group and epilepsy＋Anisomycin＋BDNF group. The immunofluorescent technique was used to identificate the hippocampal neurons. Epileptiform discharges were detected by electrophysiological techniques. Western blot assay was used to deter-mine the protein expression of TrkB and phosphorylated TrkB （p-TrkB） in all cell groups. Results （1） In ALLN group, the gray value of p-TrkB/TrkB was higher in control＋BDNF group compared with that of control group, the value was higher in epilepsy＋BDNF group than that of epilepsy group but was lower than that of control＋BDNF group. The gray value of p-TrkB/TrkB was lower in epilepsy＋ALLN＋BDNF group than that of epilepsy＋BDNF group, but no significant difference compared with that of epilepsy＋ALLN group. （2） In Anisomycin group：the gray value of p-TrkB/TrkB was higher in control＋BDNF group than that of control group. The gray value of p-TrkB/TrkB was higher in epilepsy＋BDNF group than that of epilepsy group, but which was lower than that of control＋BDNF group. The gray value of p-TrkB/TrkB was higher in epilepsy＋Aniso-mycin＋BDNF group than that of epilepsy＋BDNF group and epilepsy＋Anisomycin group. Conclusion The decreased ex-pression of TrkB.T can improve the inhibition of BDNF/TrkB signaling, and BDNF can activate BDNF/TrkB s

关 键 词：癫癎 颞叶 脑源性神经营养因子 茴香霉素 海马 神经元 钙调蛋白抑制剂 

分 类 号：R742.1[医药卫生—神经病学与精神病学]