【摘要】 为了探讨雌激素促纤溶、抗凝血的心血管保护作用及其细胞和分子生物学机制，应用全自动血凝／纤溶分析仪和发色底物显色测定、组织和细胞原位杂交及激光共聚焦扫描显微镜等技术，观察去卵巢大鼠纤维蛋白原、组织型纤溶酶原激活物和纤溶酶原激活物抑制剂－１活性及基因表达，以及补充雌激素的影响。同时观察人脐静脉内皮细胞中雌激素对组织型纤溶酶原激活物和纤溶酶原激活物抑制剂－１基因表达和活性的影响以及单个内皮细胞胞内游离钙离子的变化。结果发现，去卵巢大鼠血浆纤维蛋白原含量和纤溶酶原激活物抑制剂－１活性高于对照组（Ｐ＜０．０５，Ｐ＜０．０１），血浆组织型纤溶酶原激活物活性低于对照组，补充雌激素后，血浆纤维蛋白原含量和组织型纤溶酶原激活物活性接近对照组。去卵巢大鼠冠状动脉及心肌组织中纤溶酶原激活物抑制剂－１ｍＲＮＡ明显增高。还发现，当雌激素浓度为１０－８和１０－６ｍｏｌ／Ｌ时内皮细胞培养液中组织型纤溶酶原激活物活性高于对照组（Ｐ＜０．０５）；内皮细胞中组织型纤溶酶原激活物ｍＲＮＡ水平随雌激素浓度而升高；而纤溶酶原激活物抑制剂－１ｍＲＮＡ水平在各组中无统计学差异；内皮细胞胞内游离钙离子浓度在雌激素浓度为１０－８ｍｏｌ／Ｌ时降低，１０－６ｍ?更多还原

【Abstract】 Aim To study the effect and mechanism of cardiovascular protection of 17 beta estrodial on coagulative and fibrinolytic system in animal model and cultured human umbilical vein endothelial cell. Methods The activity changes of tissue plasminogen activator (t PA) and plasminogen activator inhibitor (PAI 1) both in vivo and in vitro were measured by spectrophotometric assay, and the level of message RNA of them were examined by in situ hybridization. In addition, the level of fibrinogen in plasma was investigated by the instrument of automobile analysing coagulative and fibrinolytic system. The intracellular free calcium concentration in single human umbilical vein endothelial cell (hUVEC) was also determined by kinetic observation with Laster Confocal Microscope. Results In vivo, the t PA activity was decreased significantly in the group that the rats were ovariectomized for 5 weeks (OVX2 group) (P<0.05) and PAI 1 activity was increased significantly in both OVX1 (the rats ovariectomized for 3 weeks) and OVX2. Fibrinogen of plasma in OVX1 was also increased (P<0.05 vs Control). In the two groups of OVX treated with 17 beta estrodial, PAI 1 activity was decreased (P<0.01, respectively) and t PA activity was increased in OVX+17 beta estrodial (P<0.05). In situ hybridization showed PAI 1 mRNA in OVX1 tissue was higher than the control. In vitro, the t PA activity and gene expression were increased with the raising concentration of 17 beta estrodial. PAI 1 mRNA was changeless between different concentrations of 17beta estrodial. The concentration of intracellular calcium free of HUVEC was decreased in physical 17 beta estrodial. Conclusions The effect of cardiovascular protect of estrogen was associated with decrease in PAI 1, fibrinogen, and [Ca 2+ ]i , as well as increase in t PA activity and gene expression.更多还原