应用荧光偏振技术研究缺氧再灌注培养血管内皮细胞膜流动性

目的　定量研究缺氧再灌注状态下血管内皮细胞膜流动性。方法　采用荧光偏振法对缺氧再灌注动物模型的培养血管内皮细胞的荧光特征光谱进行对比实验分析 ,并定量观测了缺氧再灌注对培养血管内皮细胞膜流动性的影响以及FDP和CAP的保护功能。结果　缺氧和再灌注不同时段内 ,DPH标记的血管内皮细胞的荧光光谱没有实质性改变 ,荧光强度的曲线斜率变化与对照组趋于一致 ,缺氧再灌注损伤可导致血管内皮细胞膜的荧光各向异性值降低 ,FDP和CAP对缺氧状态的培养血管内皮细胞有显著的保护作用。结论　荧光偏振技术是一种高灵敏和稳定的定量研究血管内皮细胞膜流动性的方法。

Fluorescence polarization techniques for the determination of membrane fluidity of cultured vascular endothelial cell in anoxia and reperfusion injury

ObjectiveTo perform the quantitative analysis of the membrane fluidity of vascular endothelial cell under the state of the an oxia and reperfusion.MethodsThe characteristic fluorescence spectrum of the cul tured vascular endothelial cell were comparatively analysed in the animal model of the anoxiareperfusion injury using the fluorescence polarization method . We quantitatively observed the effects of the anoxia and reperfusion on the me mbrane fluidity of cultured vascular endothelial cell and protection function of FDP and CAP.ResultsThe fluorescent spectrum of the vascular endothelial ce ll labeled by DPH did not change essentially in different times. The change of t he curve slope of the fluorescence intensity showed consistency between experime ntal and control groups. The anoxia and reperfusion injury could reduce fluoresc ence anisotropy FDP and CAP could significantly protect the cultured vascular en dothelial cell from the anoxia injury.ConclusionThe fluorescence polarization technique is a quantit ative analysis method with high sensitivity and stability on studying the membra ne fluidity of the cultured vascular endothelial cell.