CAJ | 학술논문

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。
목적：탐토전해마신경원중락안산격매수체B（TrkB）불동아형대뇌원성신경영양인자(BDNF)/TrkB신호통로적조공궤제。방법취원대배양7 d후적해마신경원，분위개조단백억제제（ALLN）조화번역억제제（An-isomycin）량대조。ALLN조우분위정상조、정상+BNDF조、전조、전+BDNF조、정상＋ALLN조、전+ALLN조화전+ALLN+BDNF조；Anisomycin조우분위정상조、정상+BDNF조、전조、전+BDNF조、정상＋Anisomycin조、전+Anisomycin조화전+Anisomycin+BDNF조。면역형광감정해마신경원，무미액처리제비전모형，전생리감정세포간양방전，면역인적기술검측각조TrkB화린산화TrkB（p-TrkB）단백적표체변화。결과（1）ALLN조：정상+BDNF조p-TrkB/TrkB회도치고우정상조；전+BDNF조고우전조，저우정상+BDNF조；전+ALLN+BDNF조저우전+BDNF조，여전+ALLN조차이무통계학의의。（2）Anisomycin조：정상+BDNF조p-TrkB/TrkB회도치고우정상조；전+BDNF조고우전조，저우정상+BDNF조；전+Anisomycin+BDNF조고우전+BDNF조화전+Anisomy-cin조。결론통과Anisomycin강저TrkB.T표체가개선전상태하BDNF/TrkB수억제적상태，통과ALLN승고TrkB. FL표체무법개선기억제상태。
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 signal pathway in epileptic hippocampal neurons. The increased TrkB.FL protein level by ALLN can’t improve the inhibition of BDNF/TrkB signal pathway.