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中国临床药理学与治疗学 ›› 2005, Vol. 10 ›› Issue (2): 158-161.

• 研究原著 • 上一篇    下一篇

在非洲爪蟾卵母细胞上表达大鼠肌肉乙酰胆碱受体的方法学研究

李传翔, 姚尚龙, 聂辉1, 张玉芹1, 吕斌2   

  1. 华中科技大学同济医学院附属协和医院麻醉科, 2环境医学研究所, 武汉430022, 湖北;
    1武汉科技大学生理教研室, 武汉430022, 湖北
  • 收稿日期:2004-11-12 修回日期:2005-01-05 出版日期:2005-02-06 发布日期:2020-11-18
  • 通讯作者: 姚尚龙, 男, 博士, 主任医师, 教授, 博士生导师, 研究方向:全麻药作用机理。Tel:027-85726834 E-mail:Yao-Shanglong@yahoo.com.cn
  • 作者简介:李传翔, 男, 博士, 主治医师, 研究方向:肌肉松驰药的基础与临床。

Study on methodology of ACh receptor's expression in Xenopus oocytes by messager RNA isolated from rat muscle

LI Chuan-xiang, YAO Shang-long, NIE Hui1, ZHANG Yu-qing1, LU Bing2   

  1. Department of Anesthesiology, Union Hospital, 1Institute of enviroment medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei, China;
    2Department of Physiology, Wuhan Technology University Wuhan 430022, Hubei, China
  • Received:2004-11-12 Revised:2005-01-05 Online:2005-02-06 Published:2020-11-18

摘要: 目的:建立适合于研究肌肉松驰药作用的乙酰胆碱受体的实验模型。方法:采用磁珠技术从大鼠肌肉中提取多聚腺嘌呤mRNA, 注入去膜的非洲爪蟾卵母细胞, 表达为有功能的离子通道, 并与未去膜和去膜未注射的卵母细胞比较, 利用电压钳方法记录电压依赖性的离子通道电流, 观察并分析三组电流特性。结果:三组卵母细胞的静息膜电位和钳制到-60 mV 时的补偿电流之间无显著的统计学意义, 通过去除卵母细胞的卵泡膜可以去除内源性受体的干扰, 在注射大鼠肌肉mRNA 的卵母细胞膜上记录到典型的N2 型ACh 受体的电流。结论:本实验建立的模型可作为在细胞和蛋白水平研究肌松药的方法。

关键词: 肌肉乙酰胆碱受体, 卵母细胞, mRNA, 电压钳技术, 方法学研究

Abstract: AIM: To establish an appropriate model for the study of the effects of muscle relaxant drug on the acetylcholine receptor and lay a methodological foundation for the study of the pharmacology and toxicology of muscle relaxant drugs on the level of cells and protein.METHODS: Poly (A)+mRNA isolated from the rat muscle by Promega's new magnesphere technique were microinjected into defolliculation's Xenopus oocytes to express functional ion channels.Voltage clamp technique was used for record the current of the three group's voltage-dependent ion channels.The three-group properties of current were analyzed.RESULTS: There was no statistically difference among three groups in mean resting potential and compensative current during holding at-60 mV.The inner receptor interference could be obliterated by oocyte defolliculation and the typical current of N2-Ach receptor could be recorded.CONCLUSION: The model set up by the test may lay a methodological foundation for the study of the pharmacology and toxicology of muscle relaxant drugs on the level of cell and protein.

Key words: muscle, ACh receptor, oocytes, mRNA, patch-clamp techniques, methodology

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