Differential transcriptional activation of matrix metalloproteinase-2 and membrane type-1 matrix metalloproteinase by experimental deep venous thrombosis.

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Presentation transcript:

Differential transcriptional activation of matrix metalloproteinase-2 and membrane type-1 matrix metalloproteinase by experimental deep venous thrombosis and thrombin Sia Dahi, BA, Jackie G. Lee, BA, David H. Lovett, MD, Rajabrata Sarkar, MD, PhD Journal of Vascular Surgery Volume 42, Issue 3, Pages 539-545 (September 2005) DOI: 10.1016/j.jvs.2005.04.051 Copyright © 2005 The Society for Vascular Surgery Terms and Conditions

Fig 1 Mouse model of deep venous thrombosis. A, Ligation of the infrarenal inferior vena cava (IVC) causes thrombus to develop. Arrowhead represents the location of the infrarenal ligature of the IVC. B, Representative sectioning of control and deep venous thrombosis (DVT) with hematoxylin and eosin counterstaining 8 days after IVC ligation (400× magnification). The vein wall is labeled by a single arrowhead. Journal of Vascular Surgery 2005 42, 539-545DOI: (10.1016/j.jvs.2005.04.051) Copyright © 2005 The Society for Vascular Surgery Terms and Conditions

Fig 2 Deep venous thrombosis (DVT) and matrix metalloproteinase (MMP)-2 activity and gene transcription in F8 transgenic mouse. A, Representative gelatin zymography 3 and 8 days after inferior vena cava (IVC) ligation. MMP-2 activity showed no significant difference at 3 days, but a noticeable induction in MMP-2 activity is observed at day 8. B, A schematic diagram of the F8 MMP-2 reporter mouse: 5086 bp of rat MMP-2 genomic DNA (including first two exons) with β-galactosidase reporter cassette. Arrow indicates MMP-2 transcription start site. Asterisk indicates mutated translational start site. C, β–galactosidase assay of control and DVT 3 and 8 days after ligation shows increased activity representing transcriptional activation of the MMP-2 gene; n = 8, *P < .05. D, Representative gelatin zymography of the IVC and superior vena cava of a mouse in which DVT was induced to illustrate that elevated levels of MMP-2 in DVT are not a result of a systematic inflammatory response. Journal of Vascular Surgery 2005 42, 539-545DOI: (10.1016/j.jvs.2005.04.051) Copyright © 2005 The Society for Vascular Surgery Terms and Conditions

Fig 3 Localization of deep venous thrombosis (DVT)-induced matrix metalloproteinase (MMP)-2 expression. A, Immunohistochemical staining for β-galactosidase (brown color) in control and DVT vein. Staining (single arrowheads) is evident in both the vein wall and thrombus of the DVT (400× magnification). B, Mouse immunoglobulin G was used in place of the primary antibody as control (400× magnification). The vein wall is labeled by double arrowheads, and staining is indicated by a single arrowhead. Journal of Vascular Surgery 2005 42, 539-545DOI: (10.1016/j.jvs.2005.04.051) Copyright © 2005 The Society for Vascular Surgery Terms and Conditions

Fig 4 Membrane-bound activator membrane type-1 matrix metalloproteinase (MT1-MMP) gene and protein expression is elevated in deep venous thrombosis (DVT). A, Real-time polymerase chain reaction analysis of MT1-MMP mRNA levels in control and ligated inferior vena cavae at 3 and 8 days after ligation. n = 8 each, *P < .05. B, Histologic staining with a MT1-MMP antibody shows increased expression relative to control of MT1-MMP in both the vein wall and thrombus (400× magnification). The vein wall is labeled by double arrowheads, and staining is indicated by a single arrowhead. Journal of Vascular Surgery 2005 42, 539-545DOI: (10.1016/j.jvs.2005.04.051) Copyright © 2005 The Society for Vascular Surgery Terms and Conditions

Fig 5 Thrombin induces differential transcriptional expression and activity of matrix metalloproteinases (MMP)-2 and membrane-bound activator membrane type-1 matrix metalloproteinase (MT1-MMP) in cultured endothelial cells. A, MMP-2 mRNA levels determined by real-time polymerase chain reaction after a 24-hour 10-U/mL thrombin treatment in human umbilical vascular endothelial cells (HUVEC), n = 5, *P < .05. B, MT1-MMP mRNA levels in thrombin-treated and control HUVEC, n = 5, *P < .05. C, Gelatin zymography of conditioned media and cell lysates of thrombin-treated and control HUVEC. Journal of Vascular Surgery 2005 42, 539-545DOI: (10.1016/j.jvs.2005.04.051) Copyright © 2005 The Society for Vascular Surgery Terms and Conditions