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目的建立膜片钳实验中适于记录延迟整流钾电流(IKM3)的单个豚鼠心室肌细胞快速分离方法,研究盐酸戊乙奎醚对IKM3的作用。方法单纯Ⅱ型胶原酶二步灌流法分离豚鼠心室肌细胞;膜片钳全细胞记录模式记录盐酸戊乙奎醚对IKM3的影响。结果分离的心肌细胞质量高,形态结构完整,成功记录了IKM3;0.2μmol·L-1盐酸戊乙奎醚在不同钳制电压下对IKM3有不同程度的抑制,试验电压为+50mV时,可使心室肌细胞上IKM3的电流密度由588.63±189.32PA降至72.85±41.91PA,抑制率为86.95%。结论二步灌流法分离得到的豚鼠心室肌细胞可用于IKM3的研究;盐酸戊乙奎醚能作用于心室肌细胞上的M3受体,减小IKM3的电流密度。
OBJECTIVE: To establish a rapid isolation method of single guinea pig ventricular myocytes suitable for recording delayed rectifier potassium current (IKM3) in patch clamp experiments to study the effect of penehyclidine hydrochloride on IKM3. Methods Pure guinea pig ventricular myocytes were isolated by two-step collagenase digestion method. Whole-cell recording mode was used to record the effect of penehyclidine hydrochloride on IKM3. Results The isolated cardiomyocytes were of high quality and intact morphology and successfully recorded IKM3. Penehyclidine hydrochloride (0.2μmol·L -1) inhibited IKM3 to varying degrees under different clamping voltages. When the test voltage was +50 mV, The current density of IKM3 on ventricular myocytes decreased from 588.63 ± 189.32PA to 72.85 ± 41.91PA with the inhibition rate of 86.95%. Conclusion Guinea pig ventricular myocytes isolated by two-step perfusion method can be used for IKM3 research. Penehyclidine hydrochloride can act on M3 receptors on ventricular myocytes and decrease the current density of IKM3.