Thrombin Induces the Expression of Tissue Factor in Human Aortic Endothelial Cells
Huang-Joe Wang, Huey‐Chun Huang, Yi‐Ching Chuang, Haimei Huang
Abstract
Huang-Joe Wang, Huey‐Chun Huang, Yi‐Ching Chuang, Haimei Huang
Abstract
Background: Atherosclerotic plaque disruption and subsequent thrombosis formation is responsible for many thrombotic complications in cardiovascular disease. Tissue factor (TF) exposure upon plaque rupture can trigger thrombin formation. Thrombin, on the other hand, can induce tissue factor expression in human umbilical vein endothelial cells or saphenous vein endothelial cells. The impact of thrombin in the endothelial cells of arterial side was less known. Methods: Human aortic endothelial cells (HAEC) were cultivated and methoxyphenyl tetrazolium inner salt assays were used to determine the non-toxicity of thrombin to endothelial cells. Real-time PCR was used to determine the relative TF mRNA quantity. Western blot was used to determine the relative protein level. TF functional activity was determined by a chromogenic assay. Results: Thrombin was non-toxic to the HAEC. TF mRNA relative quantity was enhanced 390 114 by thrombin (P = 0.034). TF protein relative quantity was enhanced 332 39 by thrombin (P = 0.01). TF activity was enhanced 547 54 by thrombin (P < 0.001). Conclusion: Thrombin is a sufficient stress to induce TF expression in HAEC. The transcriptional control by thrombin causes an increase in TF mRNA. This increase in mRNA is modestly paralleled by an increase in protein level and functional activity.
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Background: Atherosclerotic plaque disruption and subsequent thrombosis formation is responsible for many thrombotic complications in cardiovascular disease. Tissue factor (TF) exposure upon plaque rupture can trigger thrombin formation. Thrombin, on the other hand, can induce tissue factor expression in human umbilical vein endothelial cells or saphenous vein endothelial cells. The impact of thrombin in the endothelial cells of arterial side was less known. Methods: Human aortic endothelial cells (HAEC) were cultivated and methoxyphenyl tetrazolium inner salt assays were used to determine the non-toxicity of thrombin to endothelial cells. Real-time PCR was used to determine the relative TF mRNA quantity. Western blot was used to determine the relative protein level. TF functional activity was determined by a chromogenic assay. Results: Thrombin was non-toxic to the HAEC. TF mRNA relative quantity was enhanced 390 114 by thrombin (P = 0.034). TF protein relative quantity was enhanced 332 39 by thrombin (P = 0.01). TF activity was enhanced 547 54 by thrombin (P < 0.001). Conclusion: Thrombin is a sufficient stress to induce TF expression in HAEC. The transcriptional control by thrombin causes an increase in TF mRNA. This increase in mRNA is modestly paralleled by an increase in protein level and functional activity.
Key concepts: Thrombin, Tissue factor, Umbilical vein, Thrombomodulin, Medicine, Western blot, Thromboplastin, Molecular biology