胃肠动力药西沙比利对表达于HEK293细胞的人ether-a-go-go相关基因2通道的影响

doi:10.3969/j.issn.1006-7795.2014.06.016

首都医科大学学报 ›› 2014, Vol. 35 ›› Issue (6): 760-764.doi: 10.3969/j.issn.1006-7795.2014.06.016

• 分子细胞内分泌代谢论坛 • 上一篇下一篇

胃肠动力药西沙比利对表达于HEK293细胞的人ether-a-go-go相关基因2通道的影响

元沙沙^1,2, 黄海霞^3,4, 杨金奎^1,2

1. 首都医科大学附属北京同仁医院内分泌科, 北京 100730;
2. 糖尿病防治研究北京市重点实验室, 北京 100730;
3. 首都医科大学基础医学院生理与病理生理学系, 北京 100069;
4. 首都医科大学代谢紊乱相关心血管疾病北京市重点实验室, 北京 100069

收稿日期:2014-11-06 发布日期:2014-12-15
通讯作者: 杨金奎 E-mail:jinkui.yang@gmail.com
基金资助:
国家自然科学基金(81471014, 81270918);北京市自然科学基金(7131005)

Effects of the gastrointestinal prokinetic agent cisapride on human ether-a-go-go related gene 2 channels expressed on HEK293 cells

Yuan Shasha^1,2, Huang Haixia^3,4, Yang Jinkui^1,2

1. Department of Endocrine and Metabolic Diseases, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China;
2. Beijing Key Laboratory of Diabetes Research and Care Beijing 100730, China;
3. Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China;
4. Beijing Key Laboratory of Metabolic Disorders Related Cardiovascular Diseases, Capital Medical University, Beijing 100069, China

Received:2014-11-06 Published:2014-12-15
Supported by:
This study was supported by National Natural Science Foundation of China(81471014, 81270918); Natural Science Foundation of Beijing(7131005).

摘要/Abstract

摘要：

目的研究胃肠动力药西沙比利对于在HEK293细胞异源性表达的人ether-a-go-go相关基因2(human ether-a-go-go related gene 2,hERG2)通道性质的影响。方法用Lipofectamine 2000将hERG2(AF311913)瞬时转染至HEK293细胞,使其表达hERG2通道,利用全细胞膜片钳技术记录hERG2电流。加入西沙比利(终浓度分别为0.001,0.01,0.1,1.0,10.0 μmol/L),分别记录加药前和加药后2~8 min的hERG2电流,分析西沙比利浓度和作用时间对此通道电流的影响。结果西沙比利各浓度实验组中,hERG2时间依赖性电流和尾电流幅度均下降。在去极化电压为+20 mV时,随西沙比利浓度升高,电流抑制率逐渐增加。西沙比利的抑制作用随时间延长逐渐增强,西沙比利浓度为1 μmol/L时,电流在8 min内完全消失。结论西沙比利对HEK293细胞表达的hERG2通道的时间依赖性电流和尾电流均有抑制作用,该抑制作用具有浓度依赖性和时间依赖性,即浓度越高,时间越长,抑制效果越明显,直至电流降低至稳态或消失。

关键词: HEK293细胞, 膜片钳, 人ether-a-go-go相关基因, 延迟整流钾通道, 西沙比利

Abstract:

Objective To study the effects of the gastrointestinal prokinetic agent cisapride on human ether-a-go-go related gene 2(hERG2) channels expressed on HEK293 cells. Methods hERG2 cDNA(Gene Bank accession number AF311913) was transfected transiently into the HEK293 cells using Lipofectamine 2000. HERG2 current was recorded by a whole-cell patch clamp technique before and after disposal of cisapride(0.001, 0.01, 0.1, 1.0 and 10.0 μmol/L) at the time of 2, 4, 6 and 8 min. Results The amplitude of steady-state current and peak-tail current was reduced at different cisapride concentrations. The blockage of hERG2 by cisapride is strengthened with the increasing of drug concentration at the depolarization voltage of +20 mV. The effect of cisapride is also strengthened with time, and the hERG2 current vanished at the time of 8 min with 1 μmol/L cisapride. Conclusion Cisapride blocked the hERG2 channel expressed on the HEK293 cells, including the step current and tail current. The blocking of hERG2 by cisapride is concentration dependent and time dependent simultaneously until the tail current was reduced to steady state value or disappeared completely.

Key words: HEK293, patch-clamp, human ether-a-go-go related gene 2, delayed rectifier potassium channel, cisapride

中图分类号:

元沙沙, 黄海霞, 杨金奎. 胃肠动力药西沙比利对表达于HEK293细胞的人ether-a-go-go相关基因2通道的影响[J]. 首都医科大学学报, 2014, 35(6): 760-764.

Yuan Shasha, Huang Haixia, Yang Jinkui. Effects of the gastrointestinal prokinetic agent cisapride on human ether-a-go-go related gene 2 channels expressed on HEK293 cells[J]. Journal of Capital Medical University, 2014, 35(6): 760-764.

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胃肠动力药西沙比利对表达于HEK293细胞的人ether-a-go-go相关基因2通道的影响

Effects of the gastrointestinal prokinetic agent cisapride on human ether-a-go-go related gene 2 channels expressed on HEK293 cells

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