应用膜片钳技术检测药物对HERG通道抑制作用的实验过程及要点

发布时间：2018-05-07 00:42

本文选题：HERG通道 + 离子通道　；参考：《军事医学》2015年11期

【摘要】：目的应用膜片钳技术检测药物对人类ether-a-go-go相关基因(human ether-a-go-go-related gene,HERG)编码的钾离子通道的作用。探讨离子电流记录过程及实验要点。方法选择稳定转染并表达人源心肌HERG离子通道的HEK293细胞,通过全细胞记录方式,记录HERG电流。结果成功记录到HERG步阶电流和尾电流,步阶电流的Ⅰ-Ⅴ曲线表现为特征性的倒钟形结构,尾电流在去极化时迅速上升,在+10 m V时接近峰值。阳性药物E-4031能明显阻断HERG电流。检测的化合物A对HERG电流具有抑制效应,半数抑制浓度IC50为109μmol/L。结论体外HERG实验可直接检测药物对HERG电流的抑制作用,膜片钳的正确操作及电流记录是实验成败的关键因素。
[Abstract]:Objective to investigate the effect of patch clamp technique on potassium channel encoded by human ether-a-go-go-related gene Herg (HERG). The process of ion current recording and the main points of experiment are discussed. Methods HEK293 cells transfected stably and expressed human myocardial HERG ion channel were selected. The HERG currents were recorded by whole-cell recording. Results the step current and tail current of HERG were recorded successfully. The I-V curves of the step current showed a characteristic inverted bell structure. The tail current rose rapidly at depolarization and approached the peak value at 10 MV. The positive drug E-4031 could significantly block the HERG current. Compound A has inhibitory effect on HERG current, and the half inhibitory concentration of IC50 is 109 渭 mol 路L ~ (-1) 路L ~ (-1). Conclusion in vitro HERG assay can directly detect the inhibitory effect of drugs on HERG current. The correct operation of patch clamp and the recording of current are the key factors to the success of the experiment.
【作者单位】：军事医学科学院疾病预防控制所毒理学评价研究中心;
【基金】：国家科技重大专项资助项目(2009ZX09501-034)
【分类号】：R96

【参考文献】