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

以粒细胞巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ,１６ｎｇ／ｍｌ）长期（０．５－６ｄ）刺激小鼠腹腔渗了的巨噬细胞,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ＧＭ－ＣＳＦ诱导的瞬间失活的外向Ｋ电流,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４－ＡＰ高度敏感,胞内Ｃａ＾２＋浓度「Ｃａ＾２＋」升主可抑制其幅度,

Properties of the GM-CSF-induced outward K+ current in murine peritoneal exudate macrophages

Whole-cell patch-clamp technique was used to study the changes of ionic currents in murine peritoneal exudate macrophages (PEMs) prestimulated with granulocyte-macrophage colony-stimulating factor (GM-CSF, 16 ng/ml) for periods from 0.5 up to 6 d. The GM-CSF-treated PEMs developed a GM-CSF-induced transient inactivating outward K+ current (IA). 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 extracellular 4-AP (3 mmol/L). When Ca2+]i was increased (from pCa 8 to 6), the amplitudes of IA were depressed significantly. When the PEMs were exposed to cycloheximide (0.3 microgram/ml), a protein synthesis inhibitor, for 12 h, IA expression was completely suppressed. It was notable that the changes of the current expression, activation behavior and kinetic properties occurred during GM-CSF treatment. When PEMs were pretreated for a 2-d period, the frequency of IA expression reached a peak value (55% in a total of 27 cells), PEMs exhibited the highest density of the corresponding channel proteins, half-maximal activation of IA was most easily achieved with a value of -27.55 mV, and the time course of activation and inactivation during depolarization proceeded rapidly. However, along with continuous incubation with GM-CSF, the number of PEMs expressing IA decreased, the channel proteins were down regulated constantly, the activation curve for IA shifted to positive potentials, and the activation time and inactivation time of IA slowed down. These results indicated that GM-CSF could induce a transient inactivating outward K+ current in PEMs, which may have a close relation to the state of functional activation of macrophages primed with GM-CSF.