4℃预处理对人中性粒细胞膜电流和极性影响.pdfVIP

494 生理学报 Acta Physiologica Sinica, October 25, 2006, 58 (5): 494-499 实验技术 4 ℃预处理对人中性粒细胞膜电流和极性的影响 ** ** * 袁春华 ，蔡春青 ，邹 飞 南方医科大学公共卫生与热带医学学院，广州 510515 摘 要：对急性分离的人中性粒细胞采用4 ℃预处理是进行膜片钳实验前经常采取的步骤，但这一步骤对电生理记录结果 有何影响尚无文献报道。本实验探讨这一步骤对电生理记录过程和实验结果的影响。结果显示，4 ℃预处理可以显著提高 细胞的封接率，有利于对中性粒细胞进行电生理记录；封接率提高的原因与4 ℃预处理降低细胞的极性活动有关，但记录 到的电压依赖性钾通道全细胞电流和大电导Ca2+ 依赖性K+ 单通道电流动力学没有显著的变化。这些结果表明，4 ℃预处理 可能影响到细胞膜上与极性有关的脂膜变化，但对细胞膜上蛋白的功能影响较少。 关键词：中性粒细胞；极性；膜片钳技术 中图分类号：R 3 3 1 Effect of 4 ºC pretreatment on the membrane current and cell polarity in human neutrophils YUAN Chun-Hua**, CAI Chun-Qing**, ZOU Fei* School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou 510515, China Abstract: To investigate the role of ion channels in the coupling responses of neutrophils to extracellular stimulus, it is necessary to study the membrane ion channel activities using patch-clamp technique. However, little has been known about the ion channel activities in neutrophils due to the difficulties in forming giga-seal with pipettes because of small diameter of neutrophils and the easily developed polarization. Some studies indicated that favorable results could be achieved through pretreatment at low temperature before electro- physiological recordings. But it remains unclear whether the pretreatment affects the membrane current and why the seal rate increases after low temperature pretreatment. The purpose of this study was to investigate the effects of 4 ºC pretreatment on the membrane current and cell polarity in human neutrophils. In the experiments, human neutrophils were isolated from fresh peripheral blood of healthy volunteers and divided into two groups (room temperature group and 4 ºC pretreatment group). Voltage-dependent K+ (Kv)