CAJ | 학술논문

目的：研究海马神经元癫痫模型中转染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7 d后的细胞，分为正常组、正常+BDNF组、癫痫组、癫痫+BDNF组、正常转染miR-204组、癫痫转染miR-204组、癫痫转染miR-204+BDNF组。癫痫组用无镁液处理3 h制作癫痫模型。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；癫痫+BDNF组低于正常+BDNF组，高于癫痫组；癫痫+miR-204+BDNF组高于癫痫+BDNF组和癫痫+miR-204组。结论BDNF及miR-204可以改善BDNF/TrkB受抑制的状态，从而在癫痫疾病的缓解中可能发挥重要作用。
목적：연구해마신경원전간모형중전염miR-204후대뇌원성신경영양인자（BDNF）여TrkB통로조공작용적영향。방법취원대배양7 d후적세포，분위정상조、정상+BDNF조、전간조、전간+BDNF조、정상전염miR-204조、전간전염miR-204조、전간전염miR-204+BDNF조。전간조용무미액처리3 h제작전간모형。제비성공miR-204만병독표체재체。채용면역형광、막편겸급면역인적기술감정급관찰miR-204대BDNF/TrkB통로표체적영향。결과 TrkB단백적린산화수평：정상+BDNF조고우정상조；전간+BDNF조저우정상+BDNF조，고우전간조；전간+miR-204+BDNF조고우전간+BDNF조화전간+miR-204조。결론BDNF급miR-204가이개선BDNF/TrkB수억제적상태，종이재전간질병적완해중가능발휘중요작용。
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.