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用于离子通道实验的人工脂质膜的制备
  • ISSN号:1000-2669
  • 期刊名称:泸州医学院学报
  • 时间:2012
  • 页码:554-558
  • 分类:R331.38[医药卫生—人体生理学;医药卫生—基础医学]
  • 作者机构:[1]泸州医学院心血管医学研究所医学电生理实验室,四川泸州646000
  • 相关基金:国家自然基金资助项目(81173661); 四川高等学校科技创新重大培育项目(川教函[2010]597号)
  • 相关项目:活血化瘀中药(丹参)对平滑肌BKCa的激活与通道Ca2+功能域和电压感受器的相关性研究
中文摘要:

目的:摸索并建立制备平面双分子层脂质膜的方法。方法:将卵磷脂或不同比例的卵磷脂与胆固醇混合液作为成膜液,采用涂抹法制备平面双分子层脂质膜;实验中控制脂质的成分及周围的电解质环境,摸索最佳成膜条件;将含有两性霉素B的脂质体或α-toxin加入膜的一侧实验槽内,使其与双分子层脂质膜相融合并形成孔道,观察孔道形成中成膜质量。结果:最终确定的方法制备的平面双分子层脂质膜稳定性强,并能与含有两性霉素B的脂质体迅速融合,孔道形成过程中性质稳定。结论:涂抹法是一种比较稳定的脂质膜制备的简便有效方法,可为离子通道蛋白动力学及药理学特性的研究提供研究载体。

英文摘要:

Objective: To establish a stable method of reconstituting planar lipid bilayer membranes(PLBMs) for ionic channel studies.Methods: Phosphatidylcholine(PC) and the mixture of PC and cholesterol in different ratio were prepared firstly,and the PLBMs were reconstituted by painting;the composition of lipid and the surrounding electrolyte were artificially changed during the experiment to finally satisfy the best stability of the PLBMs.In order to test the stability during the pore forming in the PLBMs,the liposome of amphotericin and αtoxin were added to the cis-compartment of the CUP.Results: The PLBMs prepared by the method established in this study could last a long time and were steady during the pore forming.Conclusion: PLBMs can be reconstituted successfully by painting,and this method provides the basis for the further research of pharmacology and kinetics of ion channel protein.

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