【摘要】 目的通过白藜芦醇诱导线粒体通透过渡孔道(PTP)开放模型,介绍一种使用流式细胞技术检测线粒体PTP的新方法。方法提取大鼠肝脏线粒体,使用荧光探针NAO(nonylacridine orange)选择性的标记单个线粒体;TMRE标记检测线粒体膜电位的变化;流式细胞仪检测线粒体的侧向散射角(SSC)改变来反映白藜芦醇诱导的线粒体肿胀,即PTP的开放。结果 NAO标记线粒体结合流式细胞技术可以确定从细胞中分离线粒体的纯度;白藜芦醇处理线粒体后引起了TMRE荧光强度的降低以及SSC的减少,说明白藜芦醇可以诱导PTP开放,环孢霉素A(CsA)可以抑制白藜芦醇诱导的PTP开放。结论流式细胞技术可以准确的检测线粒体膜电位、线粒体的肿胀程度和PTP的开放。更多还原

【Abstract】 Objective To introduce a new method for detecting mitochondrial permeability transition pore(PTP) opening with flow cytometry using the resveratrol-inducing PTP opening model.Methods Mitochondria were isolated from rat livers and selectively labeled with nonyl acridine orange.The mitochondrial membrane potential was detected using flow cytometry with TMRE(tetramethylrhodamine,ethyl ester) labeling.PTP opening induced by resveratrol was represented by the changes of mitochondrial side-scattering(SSC) detected by flow cytometry.Results Flow cytometry was capable of defining the purity of the mitochondria isolated.The fluorescence intensities and SSC of the mitochondria were decreased after resveratrol treatment,indicating that resveratrol could induce PTP opening.Ciclosporin A inhibited resveratrol-induced PTP opening.Conclusion Flow cytometric analysis allows accurate and convenient detection of mitochondrial membrane potential,mitochondrial swelling and PTP opening.更多还原