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Deep vein thrombosis resolution is impaired in diet-induced type 2 diabetic mice  Fatiha Bouzeghrane, PhD, Xiaochun Zhang, MD, BSc, Guylaine Gevry, BSc,

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Presentation on theme: "Deep vein thrombosis resolution is impaired in diet-induced type 2 diabetic mice  Fatiha Bouzeghrane, PhD, Xiaochun Zhang, MD, BSc, Guylaine Gevry, BSc,"— Presentation transcript:

1 Deep vein thrombosis resolution is impaired in diet-induced type 2 diabetic mice 
Fatiha Bouzeghrane, PhD, Xiaochun Zhang, MD, BSc, Guylaine Gevry, BSc, Jean Raymond, MD  Journal of Vascular Surgery  Volume 48, Issue 6, Pages (December 2008) DOI: /j.jvs Copyright © Terms and Conditions

2 Fig 1 Mouse model of venous thrombosis. A, Endothelial damage by a neurosurgery vascular clip was followed by (B) an inferior vena cava (IVC) stenosis with a 5-0 Prolene suture (Ethicon, Somerville, NJ) placed alongside the vein. C, The vein was stenosed with the 4-0 silk ligature, and the 5-0 Prolene suture was removed once the stenosis was done. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

3 Fig 2 Transfemoral angiography of control and diabetic mice. An occluded inferior vena cava was present at 1 week in both (A) control mice and (B) diabetic mice. At 2 weeks (C) the controls showed blood flow within the thrombus core, whereas (D) blood flow in the diabetic mice (arrow) was mainly through side collaterals. Recanalization was less frequently effective in 2-week diabetic mice (1 of 10) compared with controls (9 of 10). *P < .01. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

4 Fig 3 Both (A) thrombus size and (B) weight were significantly greater in diabetic mice than in the controls at 1 and 2 weeks, and both were reduced from week 1 through 2 (*P < .05). C, A representative inferior vena cava thrombosed section is shown with nitric oxide synthase 3 (NOS3+) channels (arrow at higher magnification). Scale bar: 200 μm; inset, 40 μm. T, Thrombus; W, wall. D, At 2 weeks, diabetic mice had significantly fewer NOS3+ channels than the controls (n = 4, *P < .05). Error bars represent the standard error of the mean. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

5 Fig 4 Macrophage distribution assessed by CD68 immunoreactivity. CD68+ cells were detected in the thrombus periphery in both (A) control mice and (B) diabetic mice 1 week after surgery; and at 2 weeks after thrombus induction, sparsely in (C) the controls whereas entirely in (D) the diabetic mice. Sections are hematoxylin-counterstained. The photomicrographs are representative of results obtained from three to five sections per mouse (n = 4). Scale bar: 200 μm. E, Macrophage content increased significantly at 1 and 2 weeks in the diabetic mice compared with controls (n = 4, *P < .05). Error bars represent the standard error of the mean. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

6 Fig 5 Changes in urokinase plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) expressions in control mice and diabetic mice. Representative immunoblots of (A) uPA and (B) PAI-1 antigen levels revealed less uPA, more PAI-1, and a (C) decreased uPA/PAI-1 ratio in the diabetic mice than in the controls (n = 6, *P < .05). Error bars represent the standard error of the mean. D-G, In histologic sections, uPA and PAI-1 immunoreactivity showed (E) less uPA labeling in the vein wall (W) and thrombus section (T) of diabetic mice compared with (D) controls. In contrast, a stronger immunostaining of PAI-1 was detected in (G) diabetic mice than in (F) controls. Sections are hematoxylin-counterstained. The photomicrographs are representative of results obtained three to five sections per mouse (n = 4). Scale bar: 200 μm; inset: 40 μm. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

7 Fig 6 Picrosirius red staining of total collagen in (A) control mice 1 week and (B) diabetic mice at 1 week and (C) 2 weeks after thrombus induction. The photomicrographs are representative of results obtained three to five sections per mouse (n = 4). Scale bar: 200 μm. D, Quantification of the intensity of staining per pixel in each inferior vena cava section showed significantly less vein wall fibrosis in diabetic mice at 1 week and a greater amount at 2 weeks compared with controls (n = 4, *P < .05). Error bars represent the standard error of the mean. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

8 Fig 7 Matrix metalloproteinase (MMP) -2 and MMP-9 activities and expressions in control and diabetic mice. A, Significant augmentation in total MMP-2 and MMP-9 activities was noted in diabetic mice at (Left panel) 1 week and (Right panel) 2 weeks. Representative zymograms showed most activity was in the latent form. B, Western immunoblot analysis demonstrated increased MMP-2 and MMP-9 levels in diabetic mice 1 and 2 weeks after deep venous thrombosis compared with controls and further indicated that MMP-2 and MMP-9 proteins were decreased in diabetic mice from week 1 through 2 (n = 6, *P < 0.05). Error bars represent the standard error of the mean. C, Representative changes in MMP-2 and MMP-9 immunoreactivities of histologic sections from (Panels b, e) control and (Panels a, c, d and f) diabetic mice revealed more MMP-2 and MMP-9+ cells in vein wall (W) and thrombus section (T) in diabetic mice than in controls. Arrows indicate specific cell types expressing MMP-2 and MMP-9: 1, Pink arrow for vascular smooth muscle cells; 2, green arrow for macrophages; 3, clear arrows for endothelial cells; 4, black arrow for adventitial fibroblast. Sections are hematoxylin counterstained. The photomicrographs are representative of results obtained from three to five sections per mouse (n = 4). Scale bar: 50 μm. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © Terms and Conditions

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