The role of urokinase plasminogen activator and plasmin activator inhibitor-1 on vein wall remodeling in experimental deep vein thrombosis

• Published in: Journal of vascular surgery Vol. 56; no. 4; pp. 1089 - 1097
• Main Authors: Baldwin, Joe F., BS, Sood, Vikram, BS, Elfline, Megan A., MS, Luke, Cathy E., LVT, Dewyer, Nicholas A., MD, Diaz, Jose A., MD, Myers, Dan D., DVM, Wakefield, Thomas, MD, Henke, Peter K., MD
• Format: Journal Article
• Language: English
• Published: New York, NY Mosby, Inc 01.10.2012; Elsevier
• Subjects: Animals; Biological and medical sciences; Blood. Blood coagulation. Reticuloendothelial system; Disease Models, Animal; Endothelium, Vascular - drug effects; Endothelium, Vascular - metabolism; Endothelium, Vascular - pathology; Fibrosis; Male; Medical sciences; Mice; Pharmacology. Drug treatments; Plasminogen Activator Inhibitor 1 - pharmacology; Serine Proteinase Inhibitors - pharmacology; Surgery; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases; Urokinase-Type Plasminogen Activator - pharmacology; Urokinase-Type Plasminogen Activator - therapeutic use; Vascular surgery: aorta, extremities, vena cava. Surgery of the lymphatic vessels; Vena Cava, Inferior - drug effects; Vena Cava, Inferior - metabolism; Vena Cava, Inferior - pathology; Venous Thrombosis - drug therapy; Venous Thrombosis - metabolism; Venous Thrombosis - pathology; Vascular disease; Plasmin inhibitor; Surgery; Deep vein thrombosis; Cardiovascular disease; Venous thrombosis; Plasminogen; Venous disease
• ISSN: 0741-5214; 1097-6809
• DOI: 10.1016/j.jvs.2012.02.054

Objective Deep vein thrombosis (DVT) resolution instigates an inflammatory response, resulting in vessel wall damage and scarring. Urokinase-plasminogen activator (uPA) and its inhibitor, plasminogen activator inhibitor-1 (PAI-1), are integral components of the fibrinolytic system, essential for venous thrombosis (VT) resolution. This study determined the vein wall response when exposed to increased and decreased plasmin activity. Methods A mouse inferior vena cava (IVC) ligation model in uPA −/− or PAI-1 −/− and their genetic wild types (B6/SvEv and C57/BL6, respectively) was used to create stasis thrombi, with tissue harvest at either 8 or 21 days. Tissue analysis included gene expression of vascular smooth muscle cells (alpha smooth muscle actin [αSMA], SM22) and endothelial marker (CD31), by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, matrix metalloproteinase (MMP)-2 and -9 activity by zymography, and vein wall collagen by picro-Sirius red histologic analysis. A P < .05 was considered significant. Results Thrombi were significantly larger in both 8-day and 21-day uPA −/− as compared with wild type (WT) and were significantly smaller in both 8-day and 21-day PAI-1 −/− as compared with WT. Correspondingly, 8-day plasmin levels were reduced in half in uPA −/− and increased three-fold in PAI-1 −/− when compared with respective WT thrombi ( P < .05; n = 5-6). The endothelial marker CD31 was elevated two-fold in PAI-1 −/− mice at 8 days, but reduced 2.5-fold at 21 days in uPA −/− as compared with WT ( P = .02; n = 5-6), suggesting less endothelial preservation. Vein wall vascular smooth muscle cell (VSMC) gene expression showed that 8-day and 21-day PAI-1 −/− mice had 2.3- and 3.8-fold more SM22 and 1.8- and 2.3-fold more αSMA expression than respective WT ( P < .05; n = 5-7), as well as 1.8-fold increased αSMA (+) cells ( P ≤ .05; n = 3-5). No significant difference in MMP-2 or -9 activity was found in the PAI-1 −/− mice compared with WT, while 5.4-fold more MMP-9 was present in 21-day WT than 21-day uPA −/− ( P = .03; n = 5). Lastly, collagen was ∼two-fold greater at 8 days in PAI-1 −/− IVC as compared with WT ( P = .03; n = 6) with no differences observed in uPA −/− mice. Conclusions In stasis DVT, plasmin activity is critical for thrombus resolution. Divergent vein wall responses occur with gain or loss of plasmin activity, and despite smaller VT, greater vein wall fibrosis was associated with lack of PAI-1.