Pressure distention compared with pharmacologic relaxation in vein grafting upregulates matrix metalloproteinase-2 and -9 Ada W.Y. Chung, PhD, Pooja.

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Date of download: 6/1/2016 Copyright © 2016 American Medical Association. All rights reserved. From: Expression of Matrix Metalloproteinases and Their.

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Pressure distention compared with pharmacologic relaxation in vein grafting upregulates matrix metalloproteinase-2 and -9 Ada W.Y. Chung, PhD, Pooja Rauniyar, BSc, Honglin Luo, MD, York N. Hsiang, MD, Cornelis van Breemen, PhD, Elena B. Okon, PhD Journal of Vascular Surgery Volume 42, Issue 4, Pages 747-756 (October 2005) DOI: 10.1016/j.jvs.2005.05.037 Copyright © 2005 The Society for Vascular Surgery Terms and Conditions

Fig 1 Representative Movat stained photomicrographs. A to D, Nondistended and distended veins with different magnification; (E and F) pharmacologically treated and distended grafts; (G) area with representative extracellular matrix composition; and (H) occluded distended graft. G, The extracellular matrix components are elastin, black; collagen, yellow; and proteoglycan, green. Smooth muscle cells are purple. Arrows indicate neointima. Representative samples from five to eight independent experiments. Journal of Vascular Surgery 2005 42, 747-756DOI: (10.1016/j.jvs.2005.05.037) Copyright © 2005 The Society for Vascular Surgery Terms and Conditions

Fig 2 Morphometric analysis and extracellular matrix composition of vein and graft segments measured from Movat stained cross-sections. A, Shows ratio of intima area to media area. B, Bar graph presents the percent area of elastin, proteoglycan, and collagen. *P < .05, n = 5 to 8. Journal of Vascular Surgery 2005 42, 747-756DOI: (10.1016/j.jvs.2005.05.037) Copyright © 2005 The Society for Vascular Surgery Terms and Conditions

Fig 3 MMP-2 and MMP-9 gene transcription and protein expression in veins and grafts subjected to pressure-distention or drug treatment. A, Representative polymerase chain reaction images show the gene transcription of MMP-2, MMP-9 and β-actin (loading control). Densitometric measurement of MMP-2 transcript (normalized to the loading control). B, Bar graphs show the percentage of active MMP-2 and MMP-9 relative to the total protein expression (active + latent form) from 10 μg of sample protein extract. Insets, Representative loading control and immunoblots of the MMP-related immunoreactivity determined from Western blotting. *P < .05, paired t test; n = 7 to 8. MMP, Matrix metalloproteinase. Journal of Vascular Surgery 2005 42, 747-756DOI: (10.1016/j.jvs.2005.05.037) Copyright © 2005 The Society for Vascular Surgery Terms and Conditions

Fig 4 Proteolytic activities of MMP-2 and MMP-9 in veins and grafts subjected to pressure distention or drug treatment. Protein extract (2 μg) was electrophoresed in 8% sodium dodecyl sulfate-polyacrylamide gel electrophoresis copolymerized with gelatin (2 mg/mL). Gelatinolytic activities were identified as transparent bands against the blue background. Representative zymogram (inset) shows activities of latent MMP-9 (92 kDa), active MMP-9 (83 kDa), latent MMP-2 (68 kDa), and active MMP-2 (62 kDa). The densitometric measurement (bar graphs) shows the activities of latent MMP-9 and latent and active MMP-2. *P < .05, n = 5 to 8. MMP, Matrix metalloproteinase. Journal of Vascular Surgery 2005 42, 747-756DOI: (10.1016/j.jvs.2005.05.037) Copyright © 2005 The Society for Vascular Surgery Terms and Conditions

Fig 5 TIMP-1 and TIMP-2 gene transcription, protein expression, and activity in veins and grafts subjected to pressure distention and drug treatment. A, Representative polymerase chain reaction images show gene transcription of TIMPs. B, Bar graphs represent the densitometric measurement of TIMPs expression in 10 μg of sample protein. Representative immunoblots (insets) show the TIMP-related immunoreactivity. C, A representative reverse zymogram shows activity of TIMPs, which was visualized as dark bands against a clear background. *P < .05, n = 5 to 8. TIMP, Tissue inhibitor of matrix metalloproteinases. Journal of Vascular Surgery 2005 42, 747-756DOI: (10.1016/j.jvs.2005.05.037) Copyright © 2005 The Society for Vascular Surgery Terms and Conditions