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。
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.