豚鼠气道平滑肌的急性分离及K_(ATP)通道单通道电流的记录

Acutely Dissociated Smooth Muscle Cells from Guinea-pig Airway and Record of Single Channel Current of ATP-sensitive Potassium Channels

目的:建立急性分离豚鼠气道平滑肌的方法,并初步分析ATP敏感钾通道(KATP通道)单通道电流的性质。方法:急性分离出豚鼠3~4级支气管,并用链霉蛋白酶E分散气道平滑肌细胞,应用膜片钳技术的内面向外式记录方法,研究气道平滑肌KATP通道单通道电学性质。结果:成功记录到电导为112.4±5.14 pS的、可被优降糖所阻断的KATP通道单通道电流。钳制电压在0^-60 mV之间,通道电流无整流现象,且未见时间依赖性失活。结论:建立了急性分离豚鼠气道平滑肌的方法,并成功记录KATP通道单通道电流,为进一步研究气道平滑肌KATP通道在呼吸道疾病中的作用提供了基础。

Objective： To establish a way of acutely dissociating airway smooth muscle cells of guinea pig,and to analysis the characteristics of single channel currents of ATP-sensitive potassium channels（KATP channels）.Methods： 3 to 4 generation bronchi of guinea pig were acutely dissociated,and Pronase E was used for dispersing smooth muscle cells.Inside-out model of patch clamp technique was used to determine the electronic characteristic of single channel current of ATP-sensitive potassium channels.Results： The KATP channels with 112.4±5.14 pS single channel conductance were successfully recorded,and the activities of these channels were blocked by glibenclamide.When the membrane depolarizations were changed between 0 to-60mV,no current rectifications were found,and the channel currents showed no time-dependent inactivation.Conclusion： The way of acutely dissociating airway smooth muscle cells of guinea pig was established,and single channel current of KATP channels was successfully recorded.This work provides a basis for further study of KATP channels in respiratory tract disease.