大鼠肺动脉平滑肌细胞的分离及电压门控性钾电流的记录方法  

Isolation and Recording of Voltage Gated Potassium Channel of Pulmonary Arterial Smooth Muscle Cells in Rat

作　　者：骆红艳[1] 唐明[1] 杜以梅[1] 刘长金[1] 唐碧[1] 席姣娅[1] 洪志刚[1] Jurgen Hescheler 

机构地区：[1]华中科技大学同济医学院基础医学院生理系 [2]Institute of Neurophysiology,University of Cologne,D-50931 Cologne,Germany

出　　处：《微循环学杂志》2005年第4期9-11,F0002,共4页Chinese Journal of Microcirculation

基　　金：国家自然科学基金资助(30200098)

摘　　要：目的介绍一种大鼠肺动脉平滑肌细胞(PASMCs)的急性分离方法并观察电压门控性钾电流电生理特性。方法应用胶原酶和木瓜蛋白酶联合消化法获得大鼠PASMCs,利用全细胞膜片钳技术记录PASMCs膜上的电压门控性钾通道(Kv)电流。结果在相差显微镜下观察大鼠PASMCs呈舒展梭形,边界清晰,有完整的细胞膜,胞浆均匀,数量多,活性好。结论用酶急性分离的大鼠PASMCs,容易进行全细胞膜片钳记录,方法简单、稳定、可靠。Objective: To introduce a simple method of acutely preparing of isolated pulmonary arterial smooth muscle cells and to study the electrophysiological characteristics of voltage-gated potassium channels.Method: Single isolated pulmonary arterial smooth muscle cells were obtained by collagenase and papain digestion method and the whole cell patch clamp recording technique was used to record the voltage-gated potassium channels.Results: The morphological of isolated cells appeared to be typical fusiform shape by phase-contrast microscope. The cell membrane was integrity and nucleus was slick. The isolating method could produce adequate cells in normal form. The cells with smooth borders were chosen for whole-cell patch clamp recording.Conclusion: The established method of isolation was simple, stable and reliable.

关 键 词：肺动脉平滑肌细胞 电压门控性 缺氧性肺血管收缩反应 记录方法 电压门控钾通道 钾电流 Pulmonary Potassium Arterial 钾离子通道 

分 类 号：R543.2[医药卫生—心血管疾病] R285.5[医药卫生—内科学]