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Wanfen Xiong, MD, PhD, Rebecca Knispel, BS, Jason Mactaggart, MD, B

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Presentation on theme: "Wanfen Xiong, MD, PhD, Rebecca Knispel, BS, Jason Mactaggart, MD, B"— Presentation transcript:

1 Effects of tissue inhibitor of metalloproteinase 2 deficiency on aneurysm formation 
Wanfen Xiong, MD, PhD, Rebecca Knispel, BS, Jason Mactaggart, MD, B. Timothy Baxter, MD  Journal of Vascular Surgery  Volume 44, Issue 5, Pages (November 2006) DOI: /j.jvs Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

2 Fig 1 Reverse-zymographic analysis of tissue inhibitor of metalloproteinases (TIMPs). Aortic proteins from wild-type (WT) and TIMP-2−/− mice were extracted and separated by electrophoresis on a 17% sodium dodecylsulfate-polyacrylamide gel electrophoresis containing 0.8% gelatin and 0.16 μg/mL matrix metalloproteinase 2. The gelatin was preserved by TIMP-1 and TIMP-2 (indicated by arrows). TIMP-2 was not expressed by the TIMP-2−/− mice. The gel is representative of three separate experiments. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

3 Fig 2 Expression of tissue inhibitor of metalloproteinase (TIMP)-2. Six weeks after 0.9% NaCl or 0.25 mol/L CaCl2 treatment, wild-type mouse aortas were harvested. Total RNA from each aorta was extracted, and TIMP-2 messenger RNA (mRNA) levels were examined by reverse transcription-polymerase chain reaction. Alpha actin was used as the internal standard. The gel is representative of three separate experiments. The TIMP-2 expression is normalized to α-actin expression. The relative levels of TIMP-2 mRNA in CaCl2-treated aorta to NaCl-treated aorta are expressed in the bar graph. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

4 Fig 3 Histologic changes in mouse aorta by trichrome staining. One example of NaCl or CaCl2 treatment of C57BL/6 wild-type (WT) controls (a and c) and C57BL/6 tissue inhibitor of metalloproteinase (TIMP)-2−/− (b and d) mice is shown. The staining is representative of three to five samples with similar results. The matrix damage is profound in the CaCl2-treated WT mice. There is also significant matrix destruction in the TIMP-2−/− mice. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

5 Fig 4 Gelatin zymographic analysis of latent and processed matrix metalloproteinase (MMP)-2 in the mouse aorta. Six weeks after 0.9% NaCl or 0.25 mol/L CaCl2 treatment, mouse aortas from C57BL/6 (wild-type; WT) and tissue inhibitor of metalloproteinase (TIMP)-2−/− mice were harvested. Aortic proteins were extracted and separated by electrophoresis on a 10% sodium dodecylsulfate-polyacrylamide gel electrophoresis containing 0.8% gelatin (b). Band intensities (gelatinolytic activities) were measured by a densitometer. The percentage of processed MMP-2 and total MMP-2 in WT (n = 4) and TIMP-2−/− (n = 4) mice was analyzed (P = .0183) (a). The gel is representative of three independent experiments with similar results. MW, Molecular weight. Journal of Vascular Surgery  , DOI: ( /j.jvs ) Copyright © 2006 The Society for Vascular Surgery Terms and Conditions

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