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目的:采用全细胞膜片钳记录技术研究Sombati癫癎细胞模型电压依赖性K+电流的变化。方法:取SD新生乳鼠双侧海马,体外原代培养海马神经元,培养至第9天的部分海马神经元经无镁液处理3h制备癫癎细胞模型,采用全细胞膜片钳技术分别记录正常组(未经无镁液处理的海马神经元)和致癎组(经无镁液处理3h后更换为正常维持培养液的海马神经元)海马神经元的电压依赖性外向K+电流。结果:致癎组外向K+电流比正常组增大,峰值电流由(596.62±79.23)pA增至(978.68±53.23)pA,两组间比较差异有统计学意义(P=0.000)。与正常组相比,致癎组的I-V曲线明显左移,I-V曲线形态无显著改变。结论:Sombati癫癎细胞模型中,电压依赖性外向K+电流显著增加可能是癫癎细胞模型癎性放电后为了保持细胞稳态的代偿性保护作用。
OBJECTIVE: To study the voltage-dependent K + current changes in Sombati epileptic cell model using whole-cell patch clamp recording technique. Methods: Hippocampal neurons of SD neonatal rats were cultured in vitro. Primary cultured hippocampal neurons were cultured in vitro. Part of hippocampal neurons cultured on day 9 were treated with magnesium sulfate for 3 hours to prepare epileptic cell models. Whole-cell patch-clamp technique was used to record normal Voltage-dependent outward K + currents of hippocampal neurons were observed in the hippocampal neurons of the hippocampal neurons without hippocampal neurons treated with magnesium-free solution and hippocampal neurons of the hippocampus treated with Mg 2+ -free fluid (3h after normal magnesium treatment). Results: The extrinsic K + current in 致 group increased more than that in normal group, and the peak current increased from (596.62 ± 79.23) pA to (978.68 ± 53.23) pA. There was significant difference between the two groups (P = 0.000). Compared with the normal group, the I-V curve of the left ventricle group shifted to the left and the shape of the I-V curve did not change significantly. CONCLUSIONS: The significant increase of voltage-dependent outward K + currents in Sombati epileptic cell model may be a compensatory protective effect for keeping cell homeostasis after epileptiform cell model.