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Peter K. Henke - One of the best experts on this subject based on the ideXlab platform.
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the role of galectin 3 and galectin 3 binding protein in venous thrombosis
Blood, 2015Co-Authors: Elise P Deroo, Thomas W. Wakefield, Peter K. Henke, Daniel D Myers, Shirley K Wrobleski, Angela E Hawley, Evelyn Shea, Ramsey K Alkhalil, Jose A DiazAbstract:Galectin-3–binding protein (gal3bp) and its receptor/ligand, galectin-3 (gal3), are secreted proteins that initiate signaling cascades in several diseases, and recent human proteomic data suggest they may play a role in venous thrombosis (VT). We hypothesized that gal3bp and gal3 may promote VT. Using a mouse stasis model of VT, we found that gal3bp and gal3 were localized on Vein Wall, red blood cells, platelets, and microparticles, whereas leukocytes expressed gal3 only. Gal3 was dramatically increased during early VT and gal3bp:gal3 colocalized in the leukocyte/endothelial cell interface, where leukocytes were partially attached to the Vein Wall. Thrombus size correlated with elevated gal3 and interleukin-6 (IL-6) Vein Wall levels. Recombinant gal3 promoted VT and increased Vein Wall IL-6 mRNA. Although recombinant gal3 restored the VT size in gal3−/− mice, it had no effect on IL6−/− mice, suggesting that gal3:gal3bp promotes VT through IL-6. Moreover, significantly fewer activated neutrophils were present in the gal3−/− Vein Walls. In a group of human patients, elevated circulating gal3bp correlated with acute VT. In conclusion, gal3bp:gal3 play a critical role in VT, likely via IL-6 and PMN-mediated thrombotic mechanisms, and may be a potential biomarker in human VT.
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Statins improve the resolution of established murine venous thrombosis: reductions in thrombus burden and Vein Wall scarring.
PloS one, 2015Co-Authors: Chase W. Kessinger, Peter K. Henke, Jin Won Kim, Brian Thompson, Jason R. Mccarthy, Tetsuya Hara, Martin Sillesen, Ronan Margey, Peter Libby, Ralph WeisslederAbstract:Despite anticoagulation therapy, up to one-half of patients with deep Vein thrombosis (DVT) will develop the post-thrombotic syndrome (PTS). Improving the long-term outcome of DVT patients at risk for PTS will therefore require new approaches. Here we investigate the effects of statins—lipid-lowering agents with anti-thrombotic and anti-inflammatory properties—in decreasing thrombus burden and decreasing Vein Wall injury, mediators of PTS, in established murine stasis and non-stasis chemical-induced venous thrombosis (N = 282 mice). Treatment of mice with daily atorvastatin or rosuvastatin significantly reduced stasis venous thrombus burden by 25% without affecting lipid levels, blood coagulation parameters, or blood cell counts. Statin-driven reductions in VT burden (thrombus mass for stasis thrombi, intravital microscopy thrombus area for non-stasis thrombi) compared similarly to the therapeutic anticoagulant effects of low molecular weight heparin. Blood from statin-treated mice showed significant reductions in platelet aggregation and clot stability. Statins additionally reduced thrombus plasminogen activator inhibitor-1 (PAI-1), tissue factor, neutrophils, myeloperoxidase, neutrophil extracellular traps (NETs), and macrophages, and these effects were most notable in the earlier timepoints after DVT formation. In addition, statins reduced DVT-induced Vein Wall scarring by 50% durably up to day 21 in stasis VT, as shown by polarized light microscopy of picrosirius red-stained Vein Wall collagen. The overall results demonstrate that statins improve VT resolution via profibrinolytic, anticoagulant, antiplatelet, and anti-Vein Wall scarring effects. Statins may therefore offer a new pharmacotherapeutic approach to improve DVT resolution and to reduce the post-thrombotic syndrome, particularly in subjects who are ineligible for anticoagulation therapy.
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P-Selectin Inhibition Therapeutically Promotes Thrombus Resolution and Prevents Vein Wall Fibrosis Better Than Enoxaparin and an Inhibitor to von Willebrand Factor
Arteriosclerosis thrombosis and vascular biology, 2015Co-Authors: Jose A Diaz, Shirley K Wrobleski, Angela E Hawley, Karen J Roelofs, Christine M. Alvarado, Nichole K. Doornbos, Patrick A. Lester, Suzan E. Lowe, Joy E. Gabriel, Peter K. HenkeAbstract:Objective—Aptamers are oligonucleotides targeting protein–protein interactions with pharmacokinetic profiles and activity reversal options. Although P-selectin and von Willebrand factor (vWF) have been implicated in the development of venous thrombosis (VT), no studies have directly compared aptamer efficacy with standard of care in VT. In this study, ARC5692, an anti-P-selectin aptamer, and ARC15105, an anti-vWF aptamer, were compared with low–molecular-weight heparin, enoxaparin, to test the efficacy of P-selectin or vWF inhibition in promoting thrombus resolution and preventing Vein Wall fibrosis, in a baboon model of VT. Approach and Results—Groups were as follows: treatment arm: animals received P-selectin or vWF aptamer inhibitors or enoxaparin (n=3 per group). Controls received no treatment (n=3). Prophylactic arm: animals received P-selectin inhibitor (n=4) or vWF inhibitor (n=3). Treatment arm: P-selectin-inhibitor demonstrated a significant improvement in Vein recanalization by magnetic resonanc...
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Low-molecular-weight heparin modulates Vein Wall fibrotic response in a plasminogen activator inhibitor 1-dependent manner.
Journal of vascular surgery. Venous and lymphatic disorders, 2014Co-Authors: Andrea T., Peter K. Henke, Diana M Farris, Jose A Diaz, Karen J Roelofs, Daniel A Lawrence, N. Ballard-lipka, Thomas W. WakefieldAbstract:Background Treatment with low-molecular-weight heparin (LMWH) favorably alters the Vein Wall response to deep venous thrombosis (DVT), although the mechanisms remain unclear. Previous studies have suggested that LMWH alters the levels of circulating plasminogen activator inhibitor 1 (PAI-1), a known mediator of fibrosis, and may improve endogenous fibrinolysis. We hypothesized that LMWH favorably alters the Vein Wall response by binding of PAI-1 and acceleration of fibrinolysis. Methods Wild-type and PAI-1 −/− mice underwent treatment with LMWH after induction of occlusive DVT. Vein Wall and plasma were harvested and analyzed by enzyme-linked immunosorbent assay, zymography, real-time polymerase chain reaction, and immunohistochemistry. Results Wild-type mice treated with LMWH exhibited diminished Vein Wall fibrosis (0.6 ± 0.6 vs 1.4 ± 0.2; P P −/− mice treated with LMWH were not similarly protected from fibrosis, despite improved thrombus resolution. Treatment with LMWH was associated with decreased intrathrombus interleukin-1β (68.6 ± 31.0 vs 223.4 ± 28.9 ρg/mg total protein; P −/− mice exhibited significantly elevated intrathrombus (257.2 ± 51.5 vs 14.3 ± 3.8 ρg/mg total protein; n = 5) and Vein Wall interleukin-13 (187.2 ± 57.6 vs 9.9 ± 1.1 ρg/mg total protein; P P Conclusions LMWH did not accelerate venous thrombosis resolution but did protect against Vein Wall fibrosis in a PAI-1-dependent manner in an occlusive DVT model. Lack of PAI-1 correlated with accelerated venous thrombosis resolution but no protection from fibrosis. PAI-1 inhibition as a treatment strategy for DVT is likely to accelerate clearance of the thrombus but may come at the expense of increased Vein Wall fibrosis.
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plasminogen activator 1 overexpression decreases experimental postthrombotic Vein Wall fibrosis by a non vitronectin dependent mechanism
Journal of Thrombosis and Haemostasis, 2014Co-Authors: Jose A Diaz, Thomas W. Wakefield, Diana M Farris, N Ballardlipka, Karen J Roelofs, Daniel A Lawrence, Peter K. HenkeAbstract:Summary Background Factors associated with postthrombotic syndrome are known clinically, but the underlying cellular processes at the Vein Wall are not well delineated. Prior work suggests that Vein Wall damage does not correlate with thrombus resolution but rather with plasminogen activator-1 (PAI-1) and matrix metalloproteinase (MMP) activity. Objective We hypothesized that PAI-1 would confer post venous thrombosis (VT) Vein Wall protection via a vitronectin (Vn)-dependent mechanism. Methods A stasis model of VT was used with harvest over 2 weeks, in wild-type, Vn−/−, and PAI-1–overexpressing mice (PAI-1 Tg). Results PAI-1 Tg mice had larger VT at 6 and 14 days, compared to controls, but Vn−/− mice had no alteration of VT resolution. Gene deletion of Vn resulted in an increase in, rather than the expected decrease in, circulating PAI-1 activity. While both Vn−/− and PAI-1 Tg had attenuated intimal fibrosis, PAI-1 Tg had significantly less Vein Wall collagen and a compensatory increase in collagen III gene expression. Both Vn−/− and PAI-1 Tg Vein Wall had less monocyte chemotactic factor-1 and fewer macrophages (F4/80), with significantly less MMP-2 activity and decreased TIMP-1 antigen. Ex vivo assessment of transforming growth factor β–mediated fibrotic response showed that PAI-1 Tg Vein Walls had increased profibrotic gene expression (collagens I and III, MMP-2, and α–smooth muscle actin) compared with controls, opposite of the in vivo response. Conclusions The absence of Vn increases circulating PAI-1, which positively modulates Vein Wall fibrosis in a dose-dependent manner. Translationally, PAI-1 elevation may decrease Vein Wall damage after deep Vein thrombosis, perhaps by decreasing macrophage-mediated activities.
Thomas W. Wakefield - One of the best experts on this subject based on the ideXlab platform.
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Deep Vein Thrombosis Exhibits Characteristic Serum and Vein Wall Metabolic Phenotypes in the Inferior Vena Cava Ligation Mouse Model.
European journal of vascular and endovascular surgery : the official journal of the European Society for Vascular Surgery, 2018Co-Authors: Yeji Sung, Thomas W. Wakefield, Jose A Diaz, Konstantina Spagou, Marina Kafeza, Michael Kyriakides, Brahman Dharmarajah, Joseph Shalhoub, Elaine Holmes, Alun H. DaviesAbstract:Objectives Deep Vein thrombosis (DVT) is a major health problem, responsible for significant morbidity and mortality. The identification of a simple and effective diagnostic biomarker of DVT remains a challenge. Metabolomics have recently emerged as a new powerful scientific tool to characterise metabolic phenotypes of complex diseases and investigate small molecules in biofluids. The aim of the study was to identify the blood and Vein Wall metabolomic signature of DVT in a murine experimental model. Methods An established inferior vena cava ligation mouse model of DVT (n=10) was used and compared with sham surgery controls (n=10). Comprehensive untargeted metabolic profiling of serum and Vein Wall extracts was undertaken using liquid chromatography coupled mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR) spectroscopy. Results Multivariate and univariate statistical analysis demonstrated a differential metabolic profile when comparing DVT mice and control animals. Serum from DVT mice was characterised by differential concentrations of adenosine (decreased in DVT mice 9.6 fold), adenine (decreased 10.6 fold), and tricyclic acid cycle (TCA) intermediates, including citrate, succinate, and fumarate (1.5, 2.3, and 2.8 fold decreases, respectively). l -carnitine was found to be of greater abundance in the serum of DVT animals (67.0 fold change). A number of lipid moiety classes, including sphingomyelins, phosphatidylcholines, and triglycerides, were differentially abundant. Several metabolites were found in Vein Wall, including acetylcarnitine (increased in DVT mice 1.9 fold), adenosine (increased 2.2 fold), and ceramide (increased 2.7 fold). Correlation analysis illustrated the biochemical relationships between assigned metabolites, with the discriminatory molecules being highly correlated with each other, in both serum and Vein Wall. Conclusions The present findings demonstrate that metabolic dysregulations in DVT centre on energy metabolism, sphingolipid, and adenosine metabolism, representing a DVT specific metabolite signature in a murine experimental model.
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the role of galectin 3 and galectin 3 binding protein in venous thrombosis
Blood, 2015Co-Authors: Elise P Deroo, Thomas W. Wakefield, Peter K. Henke, Daniel D Myers, Shirley K Wrobleski, Angela E Hawley, Evelyn Shea, Ramsey K Alkhalil, Jose A DiazAbstract:Galectin-3–binding protein (gal3bp) and its receptor/ligand, galectin-3 (gal3), are secreted proteins that initiate signaling cascades in several diseases, and recent human proteomic data suggest they may play a role in venous thrombosis (VT). We hypothesized that gal3bp and gal3 may promote VT. Using a mouse stasis model of VT, we found that gal3bp and gal3 were localized on Vein Wall, red blood cells, platelets, and microparticles, whereas leukocytes expressed gal3 only. Gal3 was dramatically increased during early VT and gal3bp:gal3 colocalized in the leukocyte/endothelial cell interface, where leukocytes were partially attached to the Vein Wall. Thrombus size correlated with elevated gal3 and interleukin-6 (IL-6) Vein Wall levels. Recombinant gal3 promoted VT and increased Vein Wall IL-6 mRNA. Although recombinant gal3 restored the VT size in gal3−/− mice, it had no effect on IL6−/− mice, suggesting that gal3:gal3bp promotes VT through IL-6. Moreover, significantly fewer activated neutrophils were present in the gal3−/− Vein Walls. In a group of human patients, elevated circulating gal3bp correlated with acute VT. In conclusion, gal3bp:gal3 play a critical role in VT, likely via IL-6 and PMN-mediated thrombotic mechanisms, and may be a potential biomarker in human VT.
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Low-molecular-weight heparin modulates Vein Wall fibrotic response in a plasminogen activator inhibitor 1-dependent manner.
Journal of vascular surgery. Venous and lymphatic disorders, 2014Co-Authors: Andrea T., Peter K. Henke, Diana M Farris, Jose A Diaz, Karen J Roelofs, Daniel A Lawrence, N. Ballard-lipka, Thomas W. WakefieldAbstract:Background Treatment with low-molecular-weight heparin (LMWH) favorably alters the Vein Wall response to deep venous thrombosis (DVT), although the mechanisms remain unclear. Previous studies have suggested that LMWH alters the levels of circulating plasminogen activator inhibitor 1 (PAI-1), a known mediator of fibrosis, and may improve endogenous fibrinolysis. We hypothesized that LMWH favorably alters the Vein Wall response by binding of PAI-1 and acceleration of fibrinolysis. Methods Wild-type and PAI-1 −/− mice underwent treatment with LMWH after induction of occlusive DVT. Vein Wall and plasma were harvested and analyzed by enzyme-linked immunosorbent assay, zymography, real-time polymerase chain reaction, and immunohistochemistry. Results Wild-type mice treated with LMWH exhibited diminished Vein Wall fibrosis (0.6 ± 0.6 vs 1.4 ± 0.2; P P −/− mice treated with LMWH were not similarly protected from fibrosis, despite improved thrombus resolution. Treatment with LMWH was associated with decreased intrathrombus interleukin-1β (68.6 ± 31.0 vs 223.4 ± 28.9 ρg/mg total protein; P −/− mice exhibited significantly elevated intrathrombus (257.2 ± 51.5 vs 14.3 ± 3.8 ρg/mg total protein; n = 5) and Vein Wall interleukin-13 (187.2 ± 57.6 vs 9.9 ± 1.1 ρg/mg total protein; P P Conclusions LMWH did not accelerate venous thrombosis resolution but did protect against Vein Wall fibrosis in a PAI-1-dependent manner in an occlusive DVT model. Lack of PAI-1 correlated with accelerated venous thrombosis resolution but no protection from fibrosis. PAI-1 inhibition as a treatment strategy for DVT is likely to accelerate clearance of the thrombus but may come at the expense of increased Vein Wall fibrosis.
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plasminogen activator 1 overexpression decreases experimental postthrombotic Vein Wall fibrosis by a non vitronectin dependent mechanism
Journal of Thrombosis and Haemostasis, 2014Co-Authors: Jose A Diaz, Thomas W. Wakefield, Diana M Farris, N Ballardlipka, Karen J Roelofs, Daniel A Lawrence, Peter K. HenkeAbstract:Summary Background Factors associated with postthrombotic syndrome are known clinically, but the underlying cellular processes at the Vein Wall are not well delineated. Prior work suggests that Vein Wall damage does not correlate with thrombus resolution but rather with plasminogen activator-1 (PAI-1) and matrix metalloproteinase (MMP) activity. Objective We hypothesized that PAI-1 would confer post venous thrombosis (VT) Vein Wall protection via a vitronectin (Vn)-dependent mechanism. Methods A stasis model of VT was used with harvest over 2 weeks, in wild-type, Vn−/−, and PAI-1–overexpressing mice (PAI-1 Tg). Results PAI-1 Tg mice had larger VT at 6 and 14 days, compared to controls, but Vn−/− mice had no alteration of VT resolution. Gene deletion of Vn resulted in an increase in, rather than the expected decrease in, circulating PAI-1 activity. While both Vn−/− and PAI-1 Tg had attenuated intimal fibrosis, PAI-1 Tg had significantly less Vein Wall collagen and a compensatory increase in collagen III gene expression. Both Vn−/− and PAI-1 Tg Vein Wall had less monocyte chemotactic factor-1 and fewer macrophages (F4/80), with significantly less MMP-2 activity and decreased TIMP-1 antigen. Ex vivo assessment of transforming growth factor β–mediated fibrotic response showed that PAI-1 Tg Vein Walls had increased profibrotic gene expression (collagens I and III, MMP-2, and α–smooth muscle actin) compared with controls, opposite of the in vivo response. Conclusions The absence of Vn increases circulating PAI-1, which positively modulates Vein Wall fibrosis in a dose-dependent manner. Translationally, PAI-1 elevation may decrease Vein Wall damage after deep Vein thrombosis, perhaps by decreasing macrophage-mediated activities.
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Matrix metalloproteinase-9 deletion is associated with decreased mid-term Vein Wall fibrosis in experimental stasis DVT
Thrombosis research, 2013Co-Authors: Kristopher B. Deatrick, Megan Elfline, Catherine E. Luke, Thomas W. Wakefield, Gilbert R. Upchurch, Vikram Sood, Andrea T., Farouc A. Jaffer, Peter K. HenkeAbstract:Introduction Post thrombotic syndrome therapy is primarily palliative, and the associated Vein Wall inflammatory mechanisms are unclear. Vein Wall fibrotic injury following deep venous thrombosis (VT) is associated with elevated matrix metalloproteinases (MMPs). Whether and by what mechanism MMP9 directly contributes to Vein Wall remodeling after VT is unknown.
Gilbert R. Upchurch - One of the best experts on this subject based on the ideXlab platform.
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Matrix metalloproteinase-9 deletion is associated with decreased mid-term Vein Wall fibrosis in experimental stasis DVT
Thrombosis research, 2013Co-Authors: Kristopher B. Deatrick, Megan Elfline, Catherine E. Luke, Thomas W. Wakefield, Gilbert R. Upchurch, Vikram Sood, Andrea T., Farouc A. Jaffer, Peter K. HenkeAbstract:Introduction Post thrombotic syndrome therapy is primarily palliative, and the associated Vein Wall inflammatory mechanisms are unclear. Vein Wall fibrotic injury following deep venous thrombosis (VT) is associated with elevated matrix metalloproteinases (MMPs). Whether and by what mechanism MMP9 directly contributes to Vein Wall remodeling after VT is unknown.
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The effect of matrix metalloproteinase 2 and matrix metalloproteinase 2/9 deletion in experimental post-thrombotic Vein Wall remodeling
Journal of vascular surgery, 2013Co-Authors: Kristopher B. Deatrick, Megan Elfline, Catherine E. Luke, Thomas W. Wakefield, Gilbert R. Upchurch, Vikram Sood, Farouc A. Jaffer, Joseph F. Baldwin, Peter K. HenkeAbstract:Background Vein Wall fibrotic injury following deep venous thrombosis (VT) is associated with elevated matrix metalloproteinases (MMPs). Whether and by what mechanism MMP2 contributes to Vein Wall remodeling after VT is unknown. Methods Stasis VT was produced by ligation of the inferior vena cava and tissue was harvested at 2, 8, and 21 days in MMP2 -/- and genetic wild type (WT) mice. Tissue analysis by immunohistochemistry, enzyme-linked immunosorbent assay, real-time polymerase chain reaction, and zymography was performed. Results Thrombus resolution was less at 8 days in MMP2 -/- compared with WT, evidenced by a 51% increase in VT size ( P P P = .013), fourfold lower procollagen III gene expression ( P P P = .03) with threefold decreased apoptosis ( P P Conclusions In stasis VT, deletion of MMP2 was associated with less midterm Vein Wall fibrosis and inflammation, despite an increase in monocytes. Consideration that VT resolution was impaired with MMP2 (and MMP2/9) deletion suggests direct inhibition will likely also require anticoagulant therapy.
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Vein Wall REMODELING AFTER DEEP Vein THROMBOSIS: DIFFERENTIAL EFFECTS OF LOW MOLECULAR WEIGHT HEPARIN AND DOXYCYCLINE
Annals of vascular surgery, 2010Co-Authors: Vikram Sood, Thomas W. Wakefield, Gilbert R. Upchurch, Cathy Luke, Erin M. Miller, Mayo Mitsuya, D.d. Myers, Peter K. HenkeAbstract:OBJECTIVE Venous thrombus resolution sets up an early intense inflammatory reaction, from which Vein Wall damage results. Tissue response to injury includes matrix metalloproteinase (MMP) activation and extracellular matrix protein turnover. This study sought to determine the effect of exogenous MMP inhibition and its potential attenuation of early Vein Wall injury.
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Vein Wall re-endothelialization after deep Vein thrombosis is improved with low-molecular-weight heparin
Journal of vascular surgery, 2008Co-Authors: Daria K. Moaveni, Thomas W. Wakefield, Gilbert R. Upchurch, Erin Lynch, Vikram Sood, Cathy Luke, Peter K. HenkeAbstract:Objective Vein Wall endothelial turnover after stasis deep Vein thrombosis (DVT) has not been well characterized. The purpose of this study was to quantify re-endothelialization after DVT and determine if low-molecular-weight heparin (LMWH) therapy affects this process. Methods Stasis DVT was generated in the rat by inferior vena cava ligation, with harvest at 1, 4, and 14 days. Immunohistologic quantification of vascular smooth muscle cells and luminal endothelialization was estimated by positive staining for α-smooth muscle actin and von Willebrand factor, respectively. In separate experiments, rats were treated either before or after DVT with subcutaneous LMWH (3 mg/kg daily) until harvesting at 4 and 14 days. The inferior vena cava was processed for histologic analysis or was processed for organ culture after the thrombus was gently removed. The Vein Wall was stimulated in vitro with interleukin-1β (1 ng/mL), and the supernatant was processed at 48 hours for nitric oxide. Cells were processed by real-time polymerase chain reaction for endothelial nitric oxide synthase, inducible nitric oxide synthase, cyclooxygenase-1 and -2, and thrombomodulin at 4 and 14 days, and collagen I and III at 14 days. Comparisons were done with analysis of variance or t test. A P Results Thrombus size peaked at 4 days, whereas luminal re-endothelialization increased over time (1 day, 11% ± 2%; 4 days, 23% ± 4%; 14 days, 64% ± 7% (+) von Willebrand factor staining; P Conclusions Venous re-endothelialization occurs progressively as the DVT resolves and can be accelerated with LMWH treatment, although this effect appears limited to the early time frame. These findings may have clinical relevance for LMWH timing and treatment compared with mechanical forms of therapy.
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PLASMIN INHIBITION INCREASES MMP-9 ACTIVITY AND DECREASES Vein Wall STIFFNESS DURING VENOUS THROMBOSIS RESOLUTION
The Journal of surgical research, 2007Co-Authors: Nicholas A. Dewyer, Catherine E. Luke, Thomas W. Wakefield, Steven L. Kunkel, Gilbert R. Upchurch, Erin Lynch, Vikram Sood, Peter K. HenkeAbstract:Introduction. Deep venous thrombosis (DVT) resolution involves the plasmin and the matrix metalloproteinase (MMP) system. This study tested the hypothesis that pharmacological inhibition of the plasmin system would impair DVT resolution and worsen Vein Wall damage. Methods. A rat model of stasis DVT by inferior vena cava (IVC) ligation was performed with intravenous control saline or aprotinin (AP; 2.8 mg/kg at operation), and harvest of thrombosed IVC at 7 days. After laser Doppler imaging, DVT were separated and weighed, and Vein Wall stiffness was assessed by tensiometry. Thrombus and Vein Wall tissue analysis included total collagen by colorimetric assay, cytokines, chemokines, and D-dimer by ELISA, urokinase-plasminogen activator (uPA), and plasminogen activator inhibitor-1 (PAI-1) by immune-blotting, MMP-2 and -9 by zymography, and neutrophil (PMN) and monocyte (ED-1) leukocytes by immunohistochemistry. Results. DVT weights were 2-fold greater in the AP-treated rats (P < 0.05), but no significant differences in thrombus perfusion, collagen, or D-dimer levels were found. Vein Wall stiffness was reduced 50% (P < 0.05), suggesting less biomechanical injury. The total Vein Wall MMP-9 was increased (P < 0.05) 5-fold in the AP group compared with controls, while MMP-2 was elevated but did not reach significance. No difference was found in Vein Wall tumor necrosis factor-alpha, tissue growth factor-beta, Vein Wall or thrombus monocytes, PMN, or uPA/PAI-1 ratio between groups. Discussion. AP inhibition of the plasmin system was associated with larger thrombi but less Vein Wall injury, but no difference in other measures of resolution, possibly because of increased Vein Wall MMP-9 activity. These data suggest an important redundant mechanism for DVT resolution.
Nicholas A. Dewyer - One of the best experts on this subject based on the ideXlab platform.
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Divergent effects of Tlr9 deletion in experimental late venous thrombosis resolution and Vein Wall injury.
Thrombosis and haemostasis, 2015Co-Authors: Nicholas A. Dewyer, Megan Elfline, Catherine E. Luke, Adriana Laser, Anthony J. Comerota, Osama M. El-sayed, Nicolai A. Kittan, Ronald M. Allen, Carson Oostra, Cory M. HogaboamAbstract:Deep-Vein thrombosis (DVT) resolves via a sterile inflammatory response. Defining the inflammatory response of DVT may allow for new therapies that do not involve anticoagulation. Previously, we have shown that Toll-like receptor 9 (Tlr9) gene deleted mice had impaired venous thrombosis (VT) resolution. Here, we further characterise the role of Tlr9 signalling and sterile inflammation in chronic VT and Vein Wall responses. First, we found a human precedent exists with Tlr9+ cells present in chronic post thrombotic intraluminal tissue. Second, in a stasis VT mouse model, endogenous danger signal mediators of uric acid, HMGB-1, and neutrophil extracellular traps marker of citrullinated histone-3 (and extracellular DNA) were greater in Tlr9-/- thrombi as compared with wild-type (WT), corresponding with larger VT at 8 and 21 days. Fewer M1 type (CCR2+) monocyte/macrophages (MO) were present in Tlr9-/- thrombi than WT controls at 8 days, suggesting an impaired inflammatory cell influx. Using bone marrow-derived monocyte (BMMO) cell culture, we found decreased fibrinolytic gene expression with exposure to several endogenous danger signals. Next, adoptive transfer of cultured Tlr9+/+ BMMO to Tlr9-/- mice normalised VT resolution at 8 days. Lastly, although the VT size was larger at 21 days in Tlr9-/- mice and correlated with decreased endothelial antigen markers, no difference in fibrosis was found. These data suggest that Tlr9 signalling in MO is critical for later VT resolution, is associated with necrosis clearance, but does not affect later Vein Wall fibrosis. These findings provide insight into the Tlr9 MO mechanisms of sterile inflammation in this disease process.
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The role of urokinase plasminogen activator and plasmin activator inhibitor-1 on Vein Wall remodeling in experimental deep Vein thrombosis.
Journal of vascular surgery, 2012Co-Authors: Joseph F. Baldwin, Megan Elfline, Thomas W. Wakefield, Nicholas A. Dewyer, Vikram Sood, Jose A Diaz, Cathy Luke, D.d. Myers, Peter K. HenkeAbstract: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 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 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 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.
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PLASMIN INHIBITION INCREASES MMP-9 ACTIVITY AND DECREASES Vein Wall STIFFNESS DURING VENOUS THROMBOSIS RESOLUTION
The Journal of surgical research, 2007Co-Authors: Nicholas A. Dewyer, Catherine E. Luke, Thomas W. Wakefield, Steven L. Kunkel, Gilbert R. Upchurch, Erin Lynch, Vikram Sood, Peter K. HenkeAbstract:Introduction. Deep venous thrombosis (DVT) resolution involves the plasmin and the matrix metalloproteinase (MMP) system. This study tested the hypothesis that pharmacological inhibition of the plasmin system would impair DVT resolution and worsen Vein Wall damage. Methods. A rat model of stasis DVT by inferior vena cava (IVC) ligation was performed with intravenous control saline or aprotinin (AP; 2.8 mg/kg at operation), and harvest of thrombosed IVC at 7 days. After laser Doppler imaging, DVT were separated and weighed, and Vein Wall stiffness was assessed by tensiometry. Thrombus and Vein Wall tissue analysis included total collagen by colorimetric assay, cytokines, chemokines, and D-dimer by ELISA, urokinase-plasminogen activator (uPA), and plasminogen activator inhibitor-1 (PAI-1) by immune-blotting, MMP-2 and -9 by zymography, and neutrophil (PMN) and monocyte (ED-1) leukocytes by immunohistochemistry. Results. DVT weights were 2-fold greater in the AP-treated rats (P < 0.05), but no significant differences in thrombus perfusion, collagen, or D-dimer levels were found. Vein Wall stiffness was reduced 50% (P < 0.05), suggesting less biomechanical injury. The total Vein Wall MMP-9 was increased (P < 0.05) 5-fold in the AP group compared with controls, while MMP-2 was elevated but did not reach significance. No difference was found in Vein Wall tumor necrosis factor-alpha, tissue growth factor-beta, Vein Wall or thrombus monocytes, PMN, or uPA/PAI-1 ratio between groups. Discussion. AP inhibition of the plasmin system was associated with larger thrombi but less Vein Wall injury, but no difference in other measures of resolution, possibly because of increased Vein Wall MMP-9 activity. These data suggest an important redundant mechanism for DVT resolution.
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Neutrophils modulate post-thrombotic Vein Wall remodeling but not thrombus neovascularization
Thrombosis and haemostasis, 2006Co-Authors: Peter K. Henke, Manu R. Varma, K. Barry Deatrick, Charles G. Pearce, Andrea J. Moore, Pasu Sukheepod, Nicholas A. Dewyer, Erin Lynch, Derek A. Dubay, Gilbert R. UpchurchAbstract:Early deep venous thrombosis (DVT) resolution is associated with neutrophil (PMN) influx. This study examined the role of PMNs in thrombus neovascularization and Vein Wall injury after DVT.A rat model of DVT by inferior vena cava (IVC) ligation was performed with control serum or rabbit anti-rat PMN serum administered perioperatively with sacrifice at 2 and 7 days.At 2 days, neutropenic rats had 1.6-fold larger thrombi (P = .04) and 1.4-fold higher femoral venous pressures by water manometry (P = .008) but no difference in thrombus neovascularization was observed. By 7 days, DVT sizes were similar, but Vein Wall injury persisted in the neutropenic rats with a 2.0-fold increase in Vein Wall stiffness by microtensiometry (P < .05), as well as a 1.2-fold increased thickness (P = .04). Collagen and profibrotic growth factors were significantly increased in neutropenic IVC at 7 days (all P < .05).Vein Wall and intrathrombus uPA byWestern immunoblotting, and intrathrombus MMP-9 gelatinase activity were significantly less in neutropenic rats than controls (P < .001). Conversely, MMP-2 was significantly elevated in neutropenic IVC at 2 days after DVT. However, neutropenia induced 24 hours after DVT formation resulted in no significant increase in Vein Wall stiffness or collagen levels at 7 days, despite 1.4-fold larger thrombi (P < .05). These data suggest a critical early role for PMN in post DVT Vein Wall remodeling.
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Vein Wall remodeling after deep Vein thrombosis involves matrix metalloproteinases and late fibrosis in a mouse model
Journal of vascular surgery, 2005Co-Authors: Kristopher B. Deatrick, Thomas W. Wakefield, Manu R. Varma, Charles G. Pearce, Andrea J. Moore, Nicholas A. Dewyer, Gilbert R. Upchurch, Erin Lynch, Jonathan L. Eliason, Peter K. HenkeAbstract:Hypothesis Deep venous thrombosis (DVT) confers Vein Wall injury associated with fibrosis and extracellular matrix (ECM) turnover, likely mediated by matrix proteases. This study investigated the expression of proteases and collagen involved in early Vein Wall remodeling. Methods In the mouse, DVT was produced by ligation of the infrarenal inferior vena cava (IVC) or sham operation, and tissue was harvested at 4, 8, and 12 days. The Vein Wall tissue was processed for real-time reverse transcriptase-polymerase chain reaction (6 to 8 per time point), Western immunoblotting (5 per time point), and gelatin zymography (5 per time point). Analysis of variance was used for multiple comparisons, and a P Results Thrombus resolution was documented by a 38% decrease in the thrombosed IVC weight from day 4 to day 12 ( P = .007). Total Vein Wall collagen increased over time, with a corresponding increase in procollagen I and III, and expression peaked at 12 days (24-fold and 6.1-fold, respectively, P P P P P Conclusions Vein Wall remodeling after DVT is similar to wound healing and is associated with increased procollagen gene expression and total collagen. It is also associated with increased early MMP-9 expression, followed by MMP-2 expression and activity after DVT resolution. Clinical Relevance Deep Vein thrombosis is an often neglected problem that long term is associated with the postphlebitic syndrome of limb swelling, pain, and often ulceration. The basic mechanisms of the Vein Wall damage that results have not been delineated. The following study describes the Vein Wall matrix metalloproteinase gene and activity response induced over time in the Vein Wall after DVT. Additionally, the corresponding collagen upregulation and proximate plasmin system mediators are determined. With this knowledge, potential therapies to reduce Vein Wall injury directly might be possible.
Kristopher B. Deatrick - One of the best experts on this subject based on the ideXlab platform.
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Matrix metalloproteinase-9 deletion is associated with decreased mid-term Vein Wall fibrosis in experimental stasis DVT
Thrombosis research, 2013Co-Authors: Kristopher B. Deatrick, Megan Elfline, Catherine E. Luke, Thomas W. Wakefield, Gilbert R. Upchurch, Vikram Sood, Andrea T., Farouc A. Jaffer, Peter K. HenkeAbstract:Introduction Post thrombotic syndrome therapy is primarily palliative, and the associated Vein Wall inflammatory mechanisms are unclear. Vein Wall fibrotic injury following deep venous thrombosis (VT) is associated with elevated matrix metalloproteinases (MMPs). Whether and by what mechanism MMP9 directly contributes to Vein Wall remodeling after VT is unknown.
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The effect of matrix metalloproteinase 2 and matrix metalloproteinase 2/9 deletion in experimental post-thrombotic Vein Wall remodeling
Journal of vascular surgery, 2013Co-Authors: Kristopher B. Deatrick, Megan Elfline, Catherine E. Luke, Thomas W. Wakefield, Gilbert R. Upchurch, Vikram Sood, Farouc A. Jaffer, Joseph F. Baldwin, Peter K. HenkeAbstract:Background Vein Wall fibrotic injury following deep venous thrombosis (VT) is associated with elevated matrix metalloproteinases (MMPs). Whether and by what mechanism MMP2 contributes to Vein Wall remodeling after VT is unknown. Methods Stasis VT was produced by ligation of the inferior vena cava and tissue was harvested at 2, 8, and 21 days in MMP2 -/- and genetic wild type (WT) mice. Tissue analysis by immunohistochemistry, enzyme-linked immunosorbent assay, real-time polymerase chain reaction, and zymography was performed. Results Thrombus resolution was less at 8 days in MMP2 -/- compared with WT, evidenced by a 51% increase in VT size ( P P P = .013), fourfold lower procollagen III gene expression ( P P P = .03) with threefold decreased apoptosis ( P P Conclusions In stasis VT, deletion of MMP2 was associated with less midterm Vein Wall fibrosis and inflammation, despite an increase in monocytes. Consideration that VT resolution was impaired with MMP2 (and MMP2/9) deletion suggests direct inhibition will likely also require anticoagulant therapy.
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Postthrombotic Vein Wall remodeling: Preliminary observations
Journal of vascular surgery, 2010Co-Authors: Kristopher B. Deatrick, Megan Elfline, Nichole Baker, Catherine E. Luke, Susan Blackburn, Catherine Stabler, Thomas W. Wakefield, Peter K. HenkeAbstract:Background Postthrombotic syndrome is characterized by a fibrotic Vein injury following deep Vein thrombosis (DVT). We sought to quantify the change in Vein Wall thickness in patients who fail to resolve DVT by 6 months and whether there were differences in blood or plasma levels of inflammatory proteins associated with venous remodeling. Methods Patients presenting with confirmed lower extremity DVT were prospectively recruited for this study. Duplex imaging of the lower extremity venous system was performed, and blood was collected at entrance and repeat evaluation with blood draw and ultrasound imaging at 1 and 6 months. DVT resolution and thickness of the Vein Wall was quantified by ultrasound imaging in each segment affected by thrombus, and a contralateral, unaffected Vein Wall served as a control. Gene and protein expression of inflammatory markers were examined from leukocytes and serum, respectively. Analysis of variance or Student t-tests were used, and a P Results Thirty-two patients (12 patients with DVT resolution at 6 months, 10 patients with persistent thrombus at 6 months, and 10 healthy controls) were compared. Both resolving and nonresolving DVT were associated with a 1.5- to 1.8-fold increased Vein Wall thickness at 6 months (P = .008) as compared with nonaffected Vein Wall segments. However, the thickness of the affected segments was 1.4-fold greater in patients who had total resolution of the DVT by 6 months than in patients who had persistent chronic thrombus 6 months after presentation (P = .01). There was a four- to five-fold increased level of matrix metalloproteinase-9 (MMP-9) antigen in thrombosed patients compared with nonthrombosed patient controls (P Conclusions This preliminary study suggests ongoing Vein Wall remodeling after DVT, measurable by ultrasound and associated with certain biomarkers. At 6 months, the Vein Wall is markedly thickened and directly correlates with resolution. This suggests that the Vein Wall response is initiated early following thrombus formation and persists even in the presence of total resolution.
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Vein Wall remodeling after deep Vein thrombosis involves matrix metalloproteinases and late fibrosis in a mouse model
Journal of vascular surgery, 2005Co-Authors: Kristopher B. Deatrick, Thomas W. Wakefield, Manu R. Varma, Charles G. Pearce, Andrea J. Moore, Nicholas A. Dewyer, Gilbert R. Upchurch, Erin Lynch, Jonathan L. Eliason, Peter K. HenkeAbstract:Hypothesis Deep venous thrombosis (DVT) confers Vein Wall injury associated with fibrosis and extracellular matrix (ECM) turnover, likely mediated by matrix proteases. This study investigated the expression of proteases and collagen involved in early Vein Wall remodeling. Methods In the mouse, DVT was produced by ligation of the infrarenal inferior vena cava (IVC) or sham operation, and tissue was harvested at 4, 8, and 12 days. The Vein Wall tissue was processed for real-time reverse transcriptase-polymerase chain reaction (6 to 8 per time point), Western immunoblotting (5 per time point), and gelatin zymography (5 per time point). Analysis of variance was used for multiple comparisons, and a P Results Thrombus resolution was documented by a 38% decrease in the thrombosed IVC weight from day 4 to day 12 ( P = .007). Total Vein Wall collagen increased over time, with a corresponding increase in procollagen I and III, and expression peaked at 12 days (24-fold and 6.1-fold, respectively, P P P P P Conclusions Vein Wall remodeling after DVT is similar to wound healing and is associated with increased procollagen gene expression and total collagen. It is also associated with increased early MMP-9 expression, followed by MMP-2 expression and activity after DVT resolution. Clinical Relevance Deep Vein thrombosis is an often neglected problem that long term is associated with the postphlebitic syndrome of limb swelling, pain, and often ulceration. The basic mechanisms of the Vein Wall damage that results have not been delineated. The following study describes the Vein Wall matrix metalloproteinase gene and activity response induced over time in the Vein Wall after DVT. Additionally, the corresponding collagen upregulation and proximate plasmin system mediators are determined. With this knowledge, potential therapies to reduce Vein Wall injury directly might be possible.
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Vein Wall remodeling after deep Vein thrombosis involves matrix metalloproteinases and late fibrosis in a mouse model
Annual Meeting of the Association for Academic Surgery, 2005Co-Authors: Kristopher B. Deatrick, Thomas W. Wakefield, Manu R. Varma, Charles G. Pearce, Andrea J. Moore, Nicholas A. Dewyer, Gilbert R. Upchurch, Erin Lynch, Jonathan L. Eliason, Peter K. HenkeAbstract:Hypothesis: Deep venous thrombosis (DVT) confers Vein Wall injury associated with fibrosis and extracellular matrix (ECM) turnover, likely mediated by matrix proteases. This study investigated the expression of proteases and collagen involved in early Vein Wall remodeling. Methods: In the mouse, DVT was produced by ligation of the infrarenal inferior vena cava (IVC) or sham operation, and tissue was harvested at 4, 8, and 12 days. The Vein Wall tissue was processed for real-time reverse transcriptase-polymerase chain reaction (6 to 8 per time point), Western immunoblotting (5 per time point), and gelatin zymography (5 per time point). Analysis of variance was used for multiple comparisons, and a P <.05 was significant. Results: Thrombus resolution was documented by a 38% decrease in the thrombosed IVC weight from day 4 to day 12 (P =.007). Total Vein Wall collagen increased over time, with a corresponding increase in procollagen I and III, and expression peaked at 12 days (24-fold and 6.1-fold, respectively, P <.02). Matrix metalloproteinase-2 (MMP-2) gene expression was 23-fold greater at 12 days after thrombus formation compared with sham or 4 days after thrombosis (P <.05). Total MMP-2 activity was also significantly elevated at 12 days compared with sham (P <.05). MMP-9 expression was 19-fold and 27-fold higher at days 4 and 8, respectively, relative to sham (P <.05), with no difference in activity. MMP-14 expression was twofold to 3.6-fold greater at day 12 compared with earlier time points and shams (P <.001), but no differences in protein levels were found. Urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) protein levels were not significantly different from sham over time; however, the ratio of uPA to PAI-1 was decreased through 8 days. Conclusions: Vein Wall remodeling after DVT is similar to wound healing and is associated with increased procollagen gene expression and total collagen. It is also associated with increased early MMP-9 expression, followed by MMP-2 expression and activity after DVT resolution. (J Vasc Surg 2005;42:140-8.) Clinical Relevance: Deep Vein thrombosis is an often neglected problem that long term is associated with the postphlebitic syndrome of limb swelling, pain, and often ulceration. The basic mechanisms of the Vein Wall damage that results have not been delineated. The following study describes the Vein Wall matrix metalloproteinase gene and activity response induced over time in the Vein Wall after DVT. Additionally, the corresponding collagen upregulation and proximate plasmin system mediators are determined. With this knowledge, potential therapies to reduce Vein Wall injury directly might be possible.
