低渗诱导人小梁细胞容积敏感性氯电流的电生理学特点

J4 ›› 2010, Vol. 36 ›› Issue (2): 242-246.

低渗诱导人小梁细胞容积敏感性氯电流的电生理学特点

王霁雪, 张文杰, 袁海波, 郑雅娟, 李桂荣

1. 吉林大学第二医院眼科|吉林长春130041；2. 吉林大学基础医学院生理学教研室|吉林长春 130021；3. 吉林大学第一医院呼吸内科|吉林长春 130021

收稿日期:2009-12-09 出版日期:2010-03-28 发布日期:2010-03-28
通讯作者: 郑雅娟（Tel：0431-88766129,E-mail：zhengyajuan124@yahoo.com.cn） E-mail:zhengyajuan124@yahoo.com.cn
基金资助:
国家自然科学基金资助课题（30571993）

Electrophysiologic characteristics of volume-sensitive chloride currents induced by extracellular hypotonicity in human trabecular meshwork cells

WANG Ji-Xue, ZHANG Wen-Jie, YUAN Hai-Bo, ZHENG Ya-Juan, LI Gui-Rong

1. Department of Ophthalmology,Second Hospital,Jilin University,Changchun 130041,China;2. Department of Physiology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China;3. Department of Respiratory Diseases,First Hospital,Jilin University,Changchun 130021,China

Received:2009-12-09 Online:2010-03-28 Published:2010-03-28
About author:王霁雪（1977-）|女|吉林省四平市人|在读医学博士|主要从事青光眼及准分子的研究。

摘要/Abstract

摘要：

目的：探讨与小梁细胞容积调节有关的氯通道电流的生理学特点，阐明该电流与容积敏感性氯通道电流的关系。方法：体外培养人小梁细胞，全细胞膜片钳记录等渗和低渗条件下小梁细胞膜上的氯通道电流，观察该电流特点。低渗状态下在浴液中加入氯离子通道阻断剂NPPB和他莫昔芬及ATP后记录电流的变化。结果：在等渗条件下，仅观察到微弱且稳定的背景电流，在+100和－100 mV电压钳钳制下，外向和内向电流密度值(pA/pF)分别为1.48±0.36 和－0.91±0.10（n=6）。低渗刺激后，细胞迅速肿胀，电流较等渗时明显增大，呈外向优势，外向和内向电流密度值(pA/pF)分别为19.94±0.87 和－6.53±0.41（n=6，P<0.01）。在低渗状态下的浴液中分别加入NPPB(100 mol/L)、他莫昔芬（50 mol/L）和ATP (2 μmol/L)20　min后，外向和内向电流密度值(pA/pF)分别为7.61±0.50和－3.70±0.25、8.21±0.63和－3.83±0.25、9.08±0.67和3.89±0.22，均较低渗时明显减小（P<0.01）。结论：细胞外低渗诱导人小梁细胞产生呈外向优势氯电流，此电流对细胞容积的改变敏感，可被NPPB、他莫昔芬和ATP抑制，该电
流符合容积敏感性氯电流的特征。

关键词: 人小梁细胞；氯化物通道；膜片钳术

Abstract:

Abstract:Objective To investigate the electrophysiologic characteristics of volume - sensitive chloride currents in human trabecular meshwork cells and clarify the relationship between this current and the volume-sensitive chloride channel.Methods Human trabecular meshwork cells were culivated in vitro,whole-cell patch-clamp was used to record the current of chloride channel of human trabecular meshwork cell membrane under the isotonic and hypotonic conditions,and the characteristics of the current and the effects of the chloride channel blockers NPPB and tamoxifen and ATP on the currents under hypotonic condition were observed.Results Under isotonic condition,only weak and stable background currents were observed;Under +100 and －100　mV voltage clamp,the outward and inward current density values（pA/pF） were 1.48±0.36 and －0.91±0.10 (n=6), respectively. The currents were increased rapidly by perfusing the cells with hypotonic solution and outward currents were prominent,the outward and inward current density values(pA/pF) were 19.94±0.87 and －0.53±0.41（n=6，P<0.01). When the solution was bathed by chloride channel blockers NPPB(100 mol/L),tamoxifen（50 mol/L）or ATP(2 μmol/L)for 20 min under hypotonic condition,the outward and inward current density values(pA/pF) were 7.61±0.50 and －3.70±0.25,8.21±0.63 and　－3.83±0.25,and 9.08±0.67 and 3.89±0.22, respectively;the outward and inward current density values were reduced significantly than those under hypotonic condition（P<0.01）.Conclusion The outward chloride currents induced by extracellular hypotonicity in human trabecular meshwork cells are prominent,which are sensitive to the change of cell volume,and can be inhibited by NPPB,tamoxifen,and ATP.The currents are in line with the characteristics of volume-sensitive chloride currents.

Key words: human trabecular meshwork cell;chloride channels;patch-clamp

中图分类号:

R331

王霁雪, 张文杰, 袁海波, 郑雅娟, 李桂荣. 低渗诱导人小梁细胞容积敏感性氯电流的电生理学特点[J]. J4, 2010, 36(2): 242-246.

WANG Ji-Xue, ZHANG Wen-Jie, YUAN Hai-Bo, ZHENG Ya-Juan, LI Gui-Rong. Electrophysiologic characteristics of volume-sensitive chloride currents induced by extracellular hypotonicity in human trabecular meshwork cells[J]. J4, 2010, 36(2): 242-246.