CAJ | 학술논문

目的：探讨“无镁细胞外液”诱导体外培养大鼠海马神经元产生癫痫样放电的可行性，以期建立难治性癫痫离体细胞模型。方法选取24 h内新生Wistar大鼠，分离海马神经元后进行原代培养，体外培养至第12天时，用“无镁细胞外液”处理3 h，应用全细胞膜片钳技术记录海马神经元的放电情况。结果在培养第12天时，神经元突起间彼此接触形成神经网络。在“无镁细胞外液”处理3h后神经元产生稳定的放电，恢复正常细胞培养液培养24 h，神经元仍可检测到自发的“癫痫样放电”。结论体外培养第12天海马神经元，在“无镁细胞外液”处理后可形成稳定的自发性癫痫样放电，为今后在细胞分子水平研究癫痫发病机制提供了一种理想模型。
목적：탐토“무미세포외액”유도체외배양대서해마신경원산생전간양방전적가행성，이기건립난치성전간리체세포모형。방법선취24 h내신생Wistar대서，분리해마신경원후진행원대배양，체외배양지제12천시，용“무미세포외액”처리3 h，응용전세포막편겸기술기록해마신경원적방전정황。결과재배양제12천시，신경원돌기간피차접촉형성신경망락。재“무미세포외액”처리3h후신경원산생은정적방전，회복정상세포배양액배양24 h，신경원잉가검측도자발적“전간양방전”。결론체외배양제12천해마신경원，재“무미세포외액”처리후가형성은정적자발성전간양방전，위금후재세포분자수평연구전간발병궤제제공료일충이상모형。
Objectvie To explore the feasibility of epileptiform discharge of rat hippocampal neurons cultured in vitro and induced by magnesium-free extracellular fluid in order to establish refractory epilepsy model in cultured cells .Mtehods The neonatal Wistar rats within 24h were used to isolate hippocampal neuron to carry out primary culture in vitro .After 12d, the hippocampal neurons were treated with magnesium free extracellular fluid for 3h,then neuronal discharge activities were recorded by whole cell patch clamping technique .Results After 12-day cell culture ,neuronal dendrites touched each other to form neural network .After 3-hour treatment by magnesium-free extracellular fluid ,the neurons produced stable discharge , however ,after 24-hour treatment with normal cell culture fluid ,the spontaneous epileptiform activity in neurons could still be observed .Conclusion The stable spontaneous epileptiform activity can be formed in hippocanlpal neurons cultured in vitro for 12d,after exposed in magnesium free extracellular fluid for 3h,which provides an ideal model for pathogenetic research about epilepsy at cell and molecular level .