穿孔膜片钳方法记录L型钙通道及脱氢紫堇碱对其影响的研究

Recording L-type calcium channel current by perforated patch clamp and effect of dehydrocorydaline on L-type calcium channel

目的比较全细胞膜片钳和穿孔膜片钳方法记录大鼠心室肌细胞L型钙通道电流随时间经过的变化差异,并观察脱氢紫堇碱对L型钙通道的影响。方法采用全细胞膜片钳和穿孔膜片钳方法记录急性分离的大鼠心室肌细胞L型钙通道电流。结果采用全细胞膜片钳法记录到的L型钙通道电流峰值在15 min内衰减了(34±23)%(n=10),采用穿孔膜片钳方法记录到的L型钙通道电流峰值15 min内仅衰减了(2.7±3.4)%(n=9);采用穿孔膜片钳方法能够记录到人参皂苷Re(100μmol.L-1)的抑制效应,而采用全细胞膜片钳方法产生的电流衰减几乎完全掩盖了人参皂苷Re的效应。脱氢紫堇碱(10,100μmol.L-1)能够抑制L型钙通道的电流峰值,抑制率分别为(9±7.5)%(n=5);(28.6±8.5)%(n=5)。结论穿孔膜片钳方法较全细胞膜片钳方法在对L型钙通道电流记录方面更具稳定性和准确性,脱氢紫堇碱能够浓度依赖性的抑制L型钙通道。

Aim To observe the difference of recording L-type calcium channel currents over time through by whole cell patch clamp and perforated patch clamp method and the effect of dehydrocorydaline on L-type calcium channel by perforated patch clamp method.Methods Whole cell patch clamp and perforated patch clamp were used to record L-type calcium channel current in acutely isolated rat ventricular myocytes.Results The L-type calcium channel current peak recorded by whole cell patch clamp decayed of the（34±23）%（n=10） within 15 minutes,while using perforated patch clamp,L-type calcium channel current peak attenuated（2.7±3.4）%（n=9） within 15 minutes;The inhibitory effect of ginsenoside Re（100 μmol·L-1） could be clearly recorded by perforated patch clamp method,while the current decaying produced by whole cell patch clamp almost completely overshadowed the effects of ginsenoside Re.Dehydrocorydaline（10,100 μmol·L-1） could inhibit L-type calcium channel current peak and the inhibitory rates were（9±7.5）%（n=5）and（28.6±8.5）%（n=5） individually.Conclusions Perforated patch clamp method has more stability and accuracy than ordinary whole-cell patch clamp technique in recording L-type calcium channel current;dehydrocorydaline can suppress L-type calcium channel in a concentration-dependent manner.