Institute of Physiology, Orbely Str. 22, 0019 Yerevan, Armenia.
Toxicon. 2012 Sep 15;60(4):607-13. doi: 10.1016/j.toxicon.2012.05.016. Epub 2012 May 31.
Viper bites is an endemic public health problem in Armenia, even in the cities. Human envenomation is often characterized by clotting disorders, hypofibrinogenemia, and local tissue necrosis. In this original study, we assess some changes of cell membranes plastic properties (namely, its microviscosity, thickness, permeability) in a rat envenomation model using the biophysical approaches. We describe the interaction of Macrovipera lebetina obtusa (MLO) venom with giant unilamellar vesicles (GUVs) composed of the native phospholipid mixtures visualized through fluorescent microscopy. GUVs with a mean diameter of 30 μm have a minimum curvature and mimic cell membranes in this respect. The membrane fluorescence probe, ANS and pyrene, were used to assess the state of membrane and specifically mark the phospholipid domains. Independent of their lipid composition, GUVs were enlarged in size as venom-dependent lipid hydrolysis proceeded. Except of the visible morphological changes, ANS and pyrene also allows us to quantify the fluidity changes in the membrane by measuring of the fluorescence intensity. The presence of viper venom in GUVs media reveals a noticeable decreasing of membrane fluidity compare the control, while the binding of fluorophores with GUVs modified by venom lead to appearance of channel activity. These studies also emphasize the importance of a membrane surface curvature for its interaction with enzymatic components of venom.
在亚美尼亚,毒蛇咬伤是一个地方性的公共卫生问题,即使在城市也是如此。人类中毒常伴有凝血功能障碍、低纤维蛋白原血症和局部组织坏死。在这项原始研究中,我们使用生物物理方法评估了一种大鼠中毒模型中细胞膜塑性特性(即微粘度、厚度和通透性)的一些变化。我们描述了 Macrovipera lebetina obtusa (MLO) 毒液与由天然磷脂混合物组成的巨大单层囊泡 (GUV) 的相互作用,通过荧光显微镜可视化。平均直径为 30 μm 的 GUV 具有最小曲率,在这方面模拟细胞膜。膜荧光探针 ANS 和芘用于评估膜状态,并特别标记磷脂域。无论其脂质组成如何,随着毒液依赖性脂质水解的进行,GUV 的大小都会增大。除了可见的形态变化外,ANS 和芘还允许我们通过测量荧光强度来定量测量膜的流动性变化。在 GUV 介质中存在蛇毒会导致与对照相比膜流动性明显降低,而荧光染料与被毒液修饰的 GUV 的结合会导致通道活性的出现。这些研究还强调了膜表面曲率对于其与毒液酶成分相互作用的重要性。