Cibenzoline, a class I antiarrhythmic agent, was investigated for its effect on the ATP-sensitive K+ channel of pancreatic beta-cells by the patch clamp technique. 2. In perforated patch clamp experiments, cibenzoline depolarized the membrane of single beta-cells and thereafter, caused firing of action potentials in the presence of 2.8 mM glucose. 3. Cibenzoline inhibited the activity of the ATP-sensitive K+ channel in cell-attached recordings in the presence of 2.8 mM glucose and evoked repetitive fluctuations of the baseline current, apparently reflecting the action potentials of the beta-cell. 4. In whole-cell clamp experiments, time-independent outward current was induced by depleting cytoplasmic ATP with 0.1 mM ATP and 0.1 mM ADP in the solution contained in the pipette. The outward current was inhibited by cibenzoline in a dose-dependent manner in the concentration range of 1 microM to 100 microM and half maximum inhibition occurred at 1.5 microM. 5. Cibenzoline blocked substantially the ATP-sensitive K+ channel current when applied at the inner side of the membrane in isolated inside-out membrane patches. 6. It is concluded that cibenzoline blocks the ATP-sensitive K+ channel of pancreatic beta-cells and, thereby, stimulates insulin secretion at sub-stimulatory levels of glucose.
摘要
苯环利定是一种I类抗心律失常药物,采用膜片钳技术研究了其对胰腺β细胞ATP敏感性钾通道的作用。2. 在穿孔膜片钳实验中,苯环利定使单个β细胞膜去极化,随后在2.8 mM葡萄糖存在的情况下引发动作电位发放。3. 在2.8 mM葡萄糖存在的情况下,苯环利定在细胞贴附式记录中抑制ATP敏感性钾通道的活性,并诱发基线电流的重复波动,这显然反映了β细胞的动作电位。4. 在全细胞膜片钳实验中,通过在移液管中的溶液中用0.1 mM ATP和0.1 mM ADP耗尽细胞质ATP来诱导时间无关的外向电流。在1 microM至100 microM的浓度范围内,苯环利定以剂量依赖性方式抑制外向电流,半数最大抑制浓度为1.5 microM。5. 当在分离的内向外膜片中于膜内侧施加时,苯环利定基本阻断了ATP敏感性钾通道电流。6. 得出的结论是,苯环利定阻断胰腺β细胞的ATP敏感性钾通道,从而在低于刺激水平的葡萄糖浓度下刺激胰岛素分泌。