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粒细胞巨噬细胞集落刺激因子诱导的巨噬细胞外向K~(+)电流的特性

张晓峰, 丰美福, 吴才宏, 周培爱

中国科学院动物研究所生物膜与膜生物工程国家重点实验室. 北京 100080；北京大学生命科学学院. 北京 100871

摘要

以粒细胞巨噬细胞集落刺激因子(GM-CSF，16ng／ml)长期(0.5～6d)刺激小鼠腹腔渗出的巨噬细胞，采用全细胞膜片箝技术研究在GM－CSF刺激过程中细胞膜电流的变化，观察到一种CM－CSF诱导的瞬间失活的外向K~(+)电流(记作I_(A))，该电流在生理电压范围内可发生稳态失活，且当施加0.5Hz的去极化脉冲刺激时其失活具有频率依赖性。该电流对胞外4AP(3mmol／L)高度敏感，胞内Ca~(2+)浓度[Ca~(2+)]；升高可抑制其幅度，环己酮亚胺(0.3μg／ml)作用细胞12h可抑制其表达。值得注意的是这种K~(+)电流的表达和相应电导的激活行为及动力学均随GM－CSF剜激时间长短而异。GM－CSF作用2d,表达该电流的细胞数增至最大(大约55％)，通道蛋白的密度亦达高峰，该电流激活的半值电导电位V1／2最低，为-27.55mV，电流激活和失活迅速。随着GM－CSF作用时间的继续延长；表达该电流的细胞数下降，通道蛋白不断下调，电流激活曲线向正电位方向移动，电流激活和失活的时间过程减慢。这些结果表明，CM-CSF可诱导一种瞬间失活的外向K~(+)电流(I_(A))的表达，提示该电流可能与GM－CSF激活的巨噬细胞的功能活性状态密切相关。

关键词： 粒细胞巨噬细胞集落刺激因子; 外向K~(+)电流; 巨噬细胞; 膜片箝

Properties of the gm-csf-induced outward K~(+) current in niurine peritoneal exudate macrophages

Zhang Xiaofeng, Feng Meifu, Wu Caihong, Zhou Peiai

National Laboratory of Biomembrane and Membrane Biotechnology Institute of Zoology，Chinese Academy of Sciences. Beijing 100080,China；College of Life Sciences，Peking University.Beijing 100871,China

Abstract

Whole-cell patch-clamp technique was used to study the changes of ionic currents in murine peritoneal exudate macrophages (PEMs ) prestimulated with granulocytemacrophage colony-stimulating factor (GM-CSF,16ng/ml) for periods from 0.5 up to 6 d. The GM-CSF-treated PEMs developed a GM-CSF-induced transient inactivating outwwh K~(+) current (I_(A)). IA showed steady-state inactivation over the physiological voltage range and possessed frequency dependence of inactivation When depolarizing pulses were applied at a frequency of 0.5 Hz.IA was selectively inhibited by extracellulax 4-AP (3 mmol/L). When [Ca~(2+)], was increased (from pCa 8 to 6), the amplitudes of IA were depressed significantly. When the PEMs were exposed to cycloheximide (0.3#mu#g/ml), a protein synthesis inhibitor, for 12 h, I_(A) expression was completely suppressed. It was notable that the changes of the current expression, activation hehavior and kinetic properties occurred during GM-CSF treatment.

Key words: Graselocyte-macrophage colony-stimulating factor;Outward K~(+) current; Macrophage;Patch-clamp

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引用本文：

张晓峰, 丰美福, 吴才宏, 周培爱. 粒细胞巨噬细胞集落刺激因子诱导的巨噬细胞外向K~(+)电流的特性 [J]. 生理学报 1998; 50 (2): .

Zhang Xiaofeng, Feng Meifu, Wu Caihong, Zhou Peiai. Properties of the gm-csf-induced outward K~(+) current in niurine peritoneal exudate macrophages. Acta Physiol Sin 1998; 50 (2): (in Chinese with English abstract).