BACKGROUND: Hyperlipidemia increases the level of blood plasminogen activator inhibitor-1 (PAI-1) that is responsible for regulating fibrinolysis by inhibiting both urokinase-type plasminogen activator (u-PA) and tissue-type plasminogen activator (t-PA). While this fibrinolytic pathway is well known, the role of PAI-1 in venous thrombosis (VT) under hyperlipidemic conditions has not been fully established. We sought to determine the effects of PAI-1 in an in vivo hyperlipidemic model of VT. METHODS: C57BL/6 wild-type (WT) mice, apolipoprotein E gene-deleted mice (ApoE-/-) having hyperlipidemia, and PAI-1 gene-deleted (PAI-1-/-) mice were used in this study. Inferior vena cava (IVC) ligation below the level of the renal veins was performed to create a stasis VT. Endpoints included measuring acute thrombosis (day 2) and chronic thrombosis (days 6 and 14). At euthanasia, blood samples were collected for plasmin and PAI-1 activity. In addition, the IVC and its thrombus were evaluated for thrombus weight (TW), u-PA activity, and differential leukocyte count while the vein wall only was analyzed for monocyte chemoattractant protein-1 (MCP-1), matrix metalloproteinase (MMP) 2, and MMP-9. RESULTS: Compared to WT at day 2, ApoE-/-mice demonstrated a statistically significant 14% increase in TW (P < .05) and a significant 41% increase in circulating PAI-1 activity (P < .05), while showing a trend of decreased plasmin activity. In addition, TW in ApoE-/-mice was 45% higher than PAI-1-/-mice at day 2 (P < .05), 33% at day 6 (P < .01), and 41% at day 14 (P < .01). ApoE-/-mice exhibited undetectable levels of u-PA in both vein wall and thrombus, compared to WT, at all time points. Also, vein wall MMP-2 was significantly decreased by 64% at day 6 (P < .01) and 58% at day 14 (P < .05). MMP-9 was significantly decreased by 71% at day 2 (P < .01) and 48% at day 6 (P < .01), in ApoE-/-mice compared to WT mice. In addition, in ApoE-/-mice, MCP-1 was significantly decreased by 38% at day 2 (P < .01) and 67% at day 6 (P < .01) vs WT mice. As expected in ApoE mice, following a decrease in MCP-1, monocyte recruitment was significantly decreased at days 6 (P < .01) and 14 (P < .05). CONCLUSIONS: A significant increase of circulating PAI-1 levels in hyperlipidemic mice correlated with an early increase in TW due to impaired fibrinolysis. The undetectable levels of u-PA in ApoE-/-mice correlated to a decrease in vein wall MMP-2, MMP-9, MCP-1, and a decrease in monocyte recruitment diminishing thrombus resolution.
BACKGROUND:Hyperlipidemia increases the level of blood plasminogen activator inhibitor-1 (PAI-1) that is responsible for regulating fibrinolysis by inhibiting both urokinase-type plasminogen activator (u-PA) and tissue-type plasminogen activator (t-PA). While this fibrinolytic pathway is well known, the role of PAI-1 in venous thrombosis (VT) under hyperlipidemic conditions has not been fully established. We sought to determine the effects of PAI-1 in an in vivo hyperlipidemic model of VT. METHODS: C57BL/6 wild-type (WT) mice, apolipoprotein E gene-deleted mice (ApoE-/-) having hyperlipidemia, and PAI-1 gene-deleted (PAI-1-/-) mice were used in this study. Inferior vena cava (IVC) ligation below the level of the renal veins was performed to create a stasis VT. Endpoints included measuring acute thrombosis (day 2) and chronic thrombosis (days 6 and 14). At euthanasia, blood samples were collected for plasmin and PAI-1 activity. In addition, the IVC and its thrombus were evaluated for thrombus weight (TW), u-PA activity, and differential leukocyte count while the vein wall only was analyzed for monocyte chemoattractant protein-1 (MCP-1), matrix metalloproteinase (MMP) 2, and MMP-9. RESULTS: Compared to WT at day 2, ApoE-/-mice demonstrated a statistically significant 14% increase in TW (P < .05) and a significant 41% increase in circulating PAI-1 activity (P < .05), while showing a trend of decreased plasmin activity. In addition, TW in ApoE-/-mice was 45% higher than PAI-1-/-mice at day 2 (P < .05), 33% at day 6 (P < .01), and 41% at day 14 (P < .01). ApoE-/-mice exhibited undetectable levels of u-PA in both vein wall and thrombus, compared to WT, at all time points. Also, vein wall MMP-2 was significantly decreased by 64% at day 6 (P < .01) and 58% at day 14 (P < .05). MMP-9 was significantly decreased by 71% at day 2 (P < .01) and 48% at day 6 (P < .01), in ApoE-/-mice compared to WT mice. In addition, in ApoE-/-mice, MCP-1 was significantly decreased by 38% at day 2 (P < .01) and 67% at day 6 (P < .01) vs WT mice. As expected in ApoEmice, following a decrease in MCP-1, monocyte recruitment was significantly decreased at days 6 (P < .01) and 14 (P < .05). CONCLUSIONS: A significant increase of circulating PAI-1 levels in hyperlipidemic mice correlated with an early increase in TW due to impaired fibrinolysis. The undetectable levels of u-PA in ApoE-/-mice correlated to a decrease in vein wall MMP-2, MMP-9, MCP-1, and a decrease in monocyte recruitment diminishing thrombus resolution.
Authors: R Seguí; A Estellés; Y Mira; F España; P Villa; C Falcó; A Vayá; S Grancha; F Ferrando; J Aznar Journal: Br J Haematol Date: 2000-10 Impact factor: 6.998
Authors: G R Upchurch; J W Ford; S J Weiss; B S Knipp; D A Peterson; R W Thompson; M J Eagleton; A J Broady; M C Proctor; J C Stanley Journal: J Vasc Surg Date: 2001-07 Impact factor: 4.268
Authors: V V Sullivan; A E Hawley; D M Farris; B S Knipp; A J Varga; S K Wrobleski; P Thanapron; M J Eagleton; D D Myers; J B Fowlkes; T W Wakefield Journal: J Surg Res Date: 2003-01 Impact factor: 2.192
Authors: Matthew J Eagleton; David A Peterson; Vita V Sullivan; Karen J Roelofs; John A Ford; James C Stanley; Gilbert R Upchurch Journal: J Surg Res Date: 2002-05-01 Impact factor: 2.192
Authors: Fabio S Lira; Jose C Rosa; Adriano E Lima-Silva; Hélio A Souza; Erico C Caperuto; Marília C Seelaender; Ana R Damaso; Lila M Oyama; Ronaldo V T Santos Journal: Diabetol Metab Syndr Date: 2010-01-22 Impact factor: 3.320
Authors: Eugene A Podrez; Tatiana V Byzova; Maria Febbraio; Robert G Salomon; Yi Ma; Manojkumar Valiyaveettil; Eugenia Poliakov; Mingjiang Sun; Paula J Finton; Brian R Curtis; Juhua Chen; Renliang Zhang; Roy L Silverstein; Stanley L Hazen Journal: Nat Med Date: 2007-08-26 Impact factor: 53.440
Authors: Arunima Ghosh; Wei Li; Maria Febbraio; Ricardo G Espinola; Keith R McCrae; Erin Cockrell; Roy L Silverstein Journal: J Clin Invest Date: 2008-05 Impact factor: 14.808
Authors: Ashley A Reinke; Shih-Hon Li; Mark Warnock; Maxim E Shaydakov; Naga Sandhya Guntaka; Enming J Su; Jose A Diaz; Cory D Emal; Daniel A Lawrence Journal: J Biol Chem Date: 2018-12-03 Impact factor: 5.157
Authors: Jose A Diaz; Andrea T Obi; Daniel D Myers; Shirley K Wrobleski; Peter K Henke; Nigel Mackman; Thomas W Wakefield Journal: Arterioscler Thromb Vasc Biol Date: 2012-03 Impact factor: 8.311
Authors: K A Patterson; X Zhang; S K Wrobleski; A E Hawley; D A Lawrence; T W Wakefield; D D Myers; J A Diaz Journal: Thromb Res Date: 2012-12-29 Impact factor: 3.944
Authors: S A Siefert; C Chabasse; S Mukhopadhyay; M H Hoofnagle; D K Strickland; R Sarkar; T M Antalis Journal: J Thromb Haemost Date: 2014-09-30 Impact factor: 5.824
Authors: Elise P DeRoo; Shirley K Wrobleski; Evelyn M Shea; Ramsey K Al-Khalil; Angela E Hawley; Peter K Henke; Daniel D Myers; Thomas W Wakefield; Jose A Diaz Journal: Blood Date: 2014-11-26 Impact factor: 22.113
Authors: Hassan Albadawi; Avery A Witting; Yash Pershad; Alex Wallace; Andrew R Fleck; Peter Hoang; Ali Khademhosseini; Rahmi Oklu Journal: Cardiovasc Diagn Ther Date: 2017-12
Authors: Sean J English; Morand R Piert; Jose A Diaz; David Gordon; Abhijit Ghosh; Louis G DʼAlecy; Steven E Whitesall; Ashish K Sharma; Elise P DeRoo; Tessa Watt; Gang Su; Peter K Henke; Jonathan L Eliason; Gorav Ailawadi; Gilbert R Upchurch Journal: Ann Surg Date: 2015-02 Impact factor: 12.969
Authors: A T Obi; J A Diaz; N L Ballard-Lipka; K J Roelofs; D M Farris; D A Lawrence; T W Wakefield; P K Henke Journal: J Thromb Haemost Date: 2014-07-23 Impact factor: 5.824