溶栓及抗凝药物对内皮细胞尿激酶型纤溶酶原激活物系统和组织因子表达的影响

Effect of pro-urokinase, rt-PA and LMWH on the expression of urokinase type plasminogen activator system and tissue factor by HPAECs

目的探讨重组尿激酶原、重组组织型纤溶酶原激活物及肝索对肺动脉内皮细胞尿激酶型纤溶酶原激活物（urokina sety peplasminogen activator，u-PA）系统和组织因子（tissue factor，TF）表达的影响。方法采用体外培养的人肺动脉内皮细胞，待细胞长满后饥饿12h，换液并加入不同药物与细胞孵育12h。根据不同的药物处理方式分为五组：正常对照组；凝血酶（1IU／m1）组；凝血酶（1IU／m1）＋重组尿激酶原（150IU／m1）组；凝血酶（1IU／m1）＋重组组织型纤溶酶原激活物（150IU／m1）组；凝血酶（1IU／m1）＋低分子肝素（10IU／m1）组。孵育结束后，收集培养液，1000r／min离心10min。ELISA法检测细胞培养液中u-PA、u-PA受体（u-PA receptor，u-PAR）、纤溶酶原激活物抑制物-1（plaminogen activator inhibitor-1，PAI-1）和TF的含量。结果凝血酶（1IU／m1）组细胞培养液中PAI-1的含量较正常对照组显著增高，u-PA、u-PAR和TF的含量较正常对照组无显著改变。重组尿激酶原组细胞培养液中u-PA和u-PAR的含量较凝血酶组显著增加，PAI-1的含量较对照组显著下降，TF的含量较对照组无显著改变。重组组织型纤溶酶原激活物组细胞培养液中U-PA的含量较凝血酶组显著增加，PAI-1的含量较对照组显著下降，TF的含量较凝血酶组显著下降。低分子肝素组细胞培养液中TF的含量均较凝血酶组及对照组明显下降，u-PA、u-PAR和PAI-1较对照组无显著改变。结论重组尿激酶原、重组组织型纤溶酶原激活物及低分子肝素都能影响人肺动脉内皮细胞的内源性纤溶和抗凝功能。重组尿激酶原能够促进肺动脉内皮细胞u-PA和u-PAR的表达和释放，抑制PAI-1的表达和释放。重组组织型纤溶酶原激活物不仅能够促进肺动脉内皮细胞u-PA的表达和释放，抑制PAI-1的表达和释放，而且能够抑制TF的表达和释放。肝素主要是抑制肺动脉内皮细胞表达和释放TF。

Objective To study the effect of pro-urokinase, recombinant tissue type plasminogen activator （rt-PA） and low molecular weight heparin （LMWH） on the expression of urokinase type plasminogen activator system and tissue factor by HPAECs. Methods When the HPAECs reached confluent, starvate for 12 h, and incubate the cells with different drugs for 12 h. Cells were classified into 5 group according to different drug .. control group, thrombin （1 IU/ml） group, thrombin （1 IU/ml） and pro-urokinase （150 IU/ml） group, thrombin （1 IU/ml） and rt-PA （150 IU/mt） group and thrombin （1 IU/ml） and LMWH （10 IU/ml） group. After incubation, collect the medium, and detect the content of wPA, u-PAR, PAI-1 and TF. Results The content of PAI-1 in thrombin （1 IU/ml） group was statistically increased compared with control group, while the content of u-PA, u-PAR, and TF did not have great change. The content of u-PA and u-PAR in pro-urokinase group was statistically increased compared with control group, the content of PAI-1 was statistically decreased, while the content of TF did not have great change. The content of u-PA in rt-PA group was statistically increased compared with control group, the content of PAI-1 and TF was statistically decreased. The content of TF in I.MWHgroup was statistically increased compared with control group, while the content of u-PA, u-PAR, and PAI-1 did not have great change. Conclusions Pro urokinase, rt-PA and LMWH have direct effect on the fibrinolytic and anti-coagulant function of HPAECs.