水蛭素抑制人脐静脉内皮细胞基质金属蛋白酶-2的生成及活化

Influence of Hirudin on the Activation of Matrix Metalloproteinase-2 in Human Umbilical Vein Endothelial Cell

研究凝血酶对人脐静脉内皮细胞表达基质金属蛋白酶的影响及重组水蛭素的作用。将原代培养人脐静脉内皮细胞(HUVEC)的第2-5代分组后与凝血酶(4.0 ku／L)共同温育,同时分别加入水蛭素(6.0 ku／L)和肝素(6.0 ku／L),在不同时间,用逆转录聚合酶链反应和免疫组织化学分析的方法评价基质金属蛋白酶-2的表达情况。结果显示凝血酶促进血管内皮细胞产生和活化基质金属蛋白酶-2。重组水蛭素可以有效地阻断凝血酶的上述作用。

To investigate whether hirudin has effects on the production and activation of matrix metallo-proteinase-2 (MMP-2) in cultured human umbilical vein endothelial cell (HUVEC) induced by thrombin. HUVECs were isolated from primary human umbilical vein. The cultured 2-5 passages were incubated with thrombin (4.0 ku/L), while hirudin (6.0 ku/L) and heparin (6.0 ku/L) were added respectively except for control group. At different time, the expression of MMP-2 was determined by RT-PCR and immunocytochemistry. It was found that thrombin could upregulate the expression of matrix MMP-2 on HUVEC with time-dependent pattern (1-48 h). Hirudin showed remarkably inhibition to activation of MMP-2.