凝血酶对大鼠脑微血管内皮细胞的影响

目的 :探讨凝血酶 (Thrombin ,TM )对脑微血管内皮的影响。方法 :将大鼠脑微血管内皮细胞进行培养 ,培养液中加入 10U的TM或10U的TM + 0 .4mU的组织蛋白酶G(CaspethsinG ,CATG ) ,相差显微镜动态观察内皮细胞形态的变化 ,免疫组织化学技术检测基质金属蛋白酶 2 (MatrixMetalloproteinase 2 ,MMP 2 )表达的改变。 结果 :TM使内皮细胞发生收缩 ,细胞收缩程度具有时间依赖性 ,使内皮细胞MMP 2表达水平明显增加。TM +CATG加入培养液后 ,细胞形态、MMP 2表达与对照组比较均无明显统计学差异 (P >0 .0 5 )。结论 :TM通过激活蛋白酶激活受体 1(proteaseactivatedreceptor 1,PAR 1) ,使内皮细胞发生收缩 ,促进MMP 2表达 ,是TM增加血脑屏障 (BloodBrainBarrier,BBB)通透性的可能机制。

Effect of Thrombin on the Cerebral Microvascular Endothelial Cell in Rats

Objective: To explore the effect of thrombin(TM) on the cerebral microvascular endothelial cell.Method: Brain microvascular endothelial cell was cultured in vitro . After TM(10 U) or TM (10 U)+ caspathsin G (CATG, 0.4 mU) were added to endothelial cell medium,the changes in the morphology of cell were dynamically observed by phase contrast light microscopy and the expression of MMP 2 protein was measured by immunohistochemistry method.Results: TM induced endothelial cell contraction in a time dependent fashion and enhanced the expression of MMP 2 protein. After TM+CATG incubation, the morphology and MMP 2 expression of cell had no significant difference compared with control group( P >0.05). Conclusion: TM induces endothelial cell contraction and promotes MMP 2 expression by activating PAR 1, which may be the mechanism of blood brain barrier opening induced by TM.