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目的:探讨凝血酶对大鼠脑微血管内皮细胞(BMVEC)的损伤以及凝血酶抑制剂水蛭素的保护作用。方法:分离新生大鼠脑微血管内皮细胞并进行培养,取第二或第三代内皮细胞进行实验。实验分成3组:对照组、凝血酶组和水蛭素组。通过透射电镜和流式细胞术检测3组细胞凋亡的情况。利用TRITC-鬼笔环肽对细胞进行染色观察3组细胞骨架的形态变化。结果:凝血酶可以增加血管内皮细胞凋亡并导致细胞骨架重排、紊乱,而水蛭素可以减轻凝血酶的这种作用。结论:凝血酶可以损害血管内皮细胞骨架,引起细胞收缩、变形,同时增加内皮细胞凋亡,破坏血脑屏障。水蛭素可以减轻这种改变。
Objective: To investigate the damage of thrombin on rat brain microvascular endothelial cells (BMVEC) and the protective effect of thrombin inhibitor hirudin. Methods: Newborn rat brain microvascular endothelial cells were isolated and cultured. The second or third generation of endothelial cells were used for experiments. The experiment was divided into 3 groups: control group, thrombin group and hirudin group. The apoptosis of the three groups was detected by transmission electron microscopy and flow cytometry. The cells were stained with TRITC-phalloidin to observe the morphological changes of the cytoskeleton in the three groups. Results: Thrombin can increase the apoptosis of vascular endothelial cells and lead to cytoskeletal rearrangements and disturbances, and hirudin can reduce this effect of thrombin. Conclusion: Thrombin can damage the vascular endothelial cell skeleton, cause cell contraction and deformation, increase apoptosis of endothelial cells and destroy blood-brain barrier. Hirudin can alleviate this change.