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Statins improve the resolution of established murine venous thrombosis: reductions in thrombus burden and vein wall scarring.

Kessinger CW, Kim JW, Henke PK, Thompson B, McCarthy JR, Hara T, Sillesen M, Margey RJ, Libby P, Weissleder R, Lin CP, Jaffer FA - PLoS ONE (2015)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Research Center, Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT
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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Statins reduce thrombus burden in non-stasis murine DVT.Thrombi were visualized on IVM via a FITC-dextran angiogram as hypointense areas in the vein (A). Thrombus area and length measurements at 20, 40, 60, and 80 μm below the superficial wall-lumen interface showed a decrease in thrombus burden in atorvastatin-treated animals vs. PBS (B, C). Representative H&E axial sections showed a decrease in luminal thrombosis area in atorvastatin-treated animals compared to PBS (D, E). Thrombus outlined in yellow (D). ATV–atorvastatin. *p<0.05; ** p<0.01. Bars represent mean±SD of n = 12 animals per group. Scale bars, 25 μm.
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pone.0116621.g002: Statins reduce thrombus burden in non-stasis murine DVT.Thrombi were visualized on IVM via a FITC-dextran angiogram as hypointense areas in the vein (A). Thrombus area and length measurements at 20, 40, 60, and 80 μm below the superficial wall-lumen interface showed a decrease in thrombus burden in atorvastatin-treated animals vs. PBS (B, C). Representative H&E axial sections showed a decrease in luminal thrombosis area in atorvastatin-treated animals compared to PBS (D, E). Thrombus outlined in yellow (D). ATV–atorvastatin. *p<0.05; ** p<0.01. Bars represent mean±SD of n = 12 animals per group. Scale bars, 25 μm.

Mentions: Non-stasis, ferric chloride-induced VT. Further experiments evaluated the effects of atorvastatin on resolution of DVT induced by topical perivenous application of ferric chloride [25–27]. In vivo structural-molecular confocal fluorescence intravital microscopy (IVM) comprehensively assessed thrombus burden, inflammation, architecture and resolution in mouse femoral DVT [27]. Confocal IVM micrographs of FITC-dextran showed thrombi as hypointense regions within the vein lumen. IVM depicted thrombus morphological features, such as area and length, at defined depths below the vein wall-lumen interface (Fig. 2A). Quantitative IVM analyses demonstrated that statin therapy reduced femoral DVT area and length at day 4 by 7–16% (12.5±3.1%, n = 12 animals per group; Fig. 2B and C). Analysis of axial histological sections of the thrombosed femoral vein further assessed thrombus burden (Fig. 2D). Statin treatment resulted in a mean 28.3% reduction in luminal thrombus burden compared to PBS-treated group (0.98±0.41 × 104 μm2 thrombus area, vs. PBS group, 1.36±0.46 × 104 μm2, p = 0.01, Fig. 2E). Compared to histological assessment, confocal IVM underestimated statin-mediated reductions in thrombus burden in murine femoral vessels, due to a light attenuation-based limitation to image the thrombus at depths below 150 μm. Accordingly, all IVM quantitative analyses used a mid-luminal thrombus volume from 20 μm to 60 μm below the superficial vein wall–lumen interface was utilized, as previously described.[27]


Statins improve the resolution of established murine venous thrombosis: reductions in thrombus burden and vein wall scarring.

Kessinger CW, Kim JW, Henke PK, Thompson B, McCarthy JR, Hara T, Sillesen M, Margey RJ, Libby P, Weissleder R, Lin CP, Jaffer FA - PLoS ONE (2015)

Statins reduce thrombus burden in non-stasis murine DVT.Thrombi were visualized on IVM via a FITC-dextran angiogram as hypointense areas in the vein (A). Thrombus area and length measurements at 20, 40, 60, and 80 μm below the superficial wall-lumen interface showed a decrease in thrombus burden in atorvastatin-treated animals vs. PBS (B, C). Representative H&E axial sections showed a decrease in luminal thrombosis area in atorvastatin-treated animals compared to PBS (D, E). Thrombus outlined in yellow (D). ATV–atorvastatin. *p<0.05; ** p<0.01. Bars represent mean±SD of n = 12 animals per group. Scale bars, 25 μm.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4334538&req=5

pone.0116621.g002: Statins reduce thrombus burden in non-stasis murine DVT.Thrombi were visualized on IVM via a FITC-dextran angiogram as hypointense areas in the vein (A). Thrombus area and length measurements at 20, 40, 60, and 80 μm below the superficial wall-lumen interface showed a decrease in thrombus burden in atorvastatin-treated animals vs. PBS (B, C). Representative H&E axial sections showed a decrease in luminal thrombosis area in atorvastatin-treated animals compared to PBS (D, E). Thrombus outlined in yellow (D). ATV–atorvastatin. *p<0.05; ** p<0.01. Bars represent mean±SD of n = 12 animals per group. Scale bars, 25 μm.
Mentions: Non-stasis, ferric chloride-induced VT. Further experiments evaluated the effects of atorvastatin on resolution of DVT induced by topical perivenous application of ferric chloride [25–27]. In vivo structural-molecular confocal fluorescence intravital microscopy (IVM) comprehensively assessed thrombus burden, inflammation, architecture and resolution in mouse femoral DVT [27]. Confocal IVM micrographs of FITC-dextran showed thrombi as hypointense regions within the vein lumen. IVM depicted thrombus morphological features, such as area and length, at defined depths below the vein wall-lumen interface (Fig. 2A). Quantitative IVM analyses demonstrated that statin therapy reduced femoral DVT area and length at day 4 by 7–16% (12.5±3.1%, n = 12 animals per group; Fig. 2B and C). Analysis of axial histological sections of the thrombosed femoral vein further assessed thrombus burden (Fig. 2D). Statin treatment resulted in a mean 28.3% reduction in luminal thrombus burden compared to PBS-treated group (0.98±0.41 × 104 μm2 thrombus area, vs. PBS group, 1.36±0.46 × 104 μm2, p = 0.01, Fig. 2E). Compared to histological assessment, confocal IVM underestimated statin-mediated reductions in thrombus burden in murine femoral vessels, due to a light attenuation-based limitation to image the thrombus at depths below 150 μm. Accordingly, all IVM quantitative analyses used a mid-luminal thrombus volume from 20 μm to 60 μm below the superficial vein wall–lumen interface was utilized, as previously described.[27]

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Research Center, Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus