Differential regulation of matrix metalloproteinase activities in abdominal aortic aneurysms Borhane Annabi, PhD, a, Daniel Shédid, MD, b, Pierre Ghosn,

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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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Presentation transcript:

Differential regulation of matrix metalloproteinase activities in abdominal aortic aneurysms Borhane Annabi, PhD, a, Daniel Shédid, MD, b, Pierre Ghosn, MD, c, Rhoda L. Kenigsberg, MD, b, Richard R. Desrosiers, PhD, a, Michel W. Bojanowski, MD, b, Édith Beaulieu, PhD, a, Edgar Nassif, MD, d, Robert Moumdjian, MD, e, Richard Béliveau, PhD, a, Journal of Vascular Surgery Volume 35, Issue 3, Pages 539-546 (March 2002) DOI: 10.1067/mva.2002.121124 Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 1 Expression of actin isoforms in aortic aneurysms. For analysis of expression of α-actin and β-actin, abdominal aortic aneurysms (AAAs; n = 4) and healthy aorta (n = 4) were homogenized. Proteins (25 mg/well) were resolved on 12.5% SDS-polyacrylamide electrophoresis gel and (A) α-actin and β-actin were immunodetected as described in Materials and Methods section. B, Quantification of α-actin (healthy aorta, 100 ± 12, versus AAA, 15 ± 4) and β-actin (healthy aorta, 100 ± 3, versus AAA, 85 ± 2) levels was performed with densitometry and is expressed as percent of values for healthy aorta. *Values of less than.05 were considered significant. NA, Healthy aorta. Journal of Vascular Surgery 2002 35, 539-546DOI: (10.1067/mva.2002.121124) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 2 Increased matrix metalloproteinase 1 (MMP-1) protein expression in aortic aneurysms. Protein homogenates of healthy aorta (n = 3) and abdominal aortic aneurysms (AAAs; n = 5) were resolved (25 mg/well) on 9% SDS-polyacrylamide electrophoresis gel as described in Materials and Methods section. A, Immunoblots of expression of active MMP-1 (45 kd) and pro–MMP-1 (53 kd) forms are shown. B, Quantification of pro–MMP-1 protein level (healthy aorta, 100 ± 22, versus AAA, 769 ± 25) was performed with densitometry and is expressed as percent of values for healthy aorta. *Values of less than.05 were considered significant. NA, Healthy aorta. Journal of Vascular Surgery 2002 35, 539-546DOI: (10.1067/mva.2002.121124) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 3 Membrane-type 1 matrix metalloproteinase (MT1-MMP) proteolytic processing is decreased in aortic aneurysms. Homogenates of healthy aorta (n = 4) and abdominal aortic aneurysms (AAAs; n = 5) were resolved (25 mg protein/well) on 9% SDS-polyacrylamide electrophoresis gel as described in Materials and Methods section. A, MT1-MMP immunoreactivity was analyzed, and 63-kd and 55-kd proteins represent unprocessed active forms of MT1-MMP, whereas 43-kd form is processed inactive form. Human recombinant MT1-MMP (rMT1-MMP) was applied on gel as standard. B, Quantification of active (healthy aorta, 100 ± 25, versus AAA, 347 ± 23) and inactive (healthy aorta, 100 ± 11, versus AAA, 22 ± 11) forms of MT1-MMP was performed with densitometry and is expressed as percent of values for healthy aorta. *Values of less than.05 were considered significant. NA, Healthy aorta. Journal of Vascular Surgery 2002 35, 539-546DOI: (10.1067/mva.2002.121124) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 4 Immunoblot analysis of expressed tissue inhibitor of matrix metalloproteinase 1 (TIMP-1), TIMP-2, and TIMP-3 in healthy aorta and aortic aneurysms. Homogenates of healthy aorta (n = 4) and abdominal aortic aneurysms (AAAs; n = 5) were resolved (25 mg protein/well) on 12.5% SDS-polyacrylamide electrophoresis gel as described in Materials and Methods section. A, Expression of TIMP-1, TIMP-2, and TIMP-3 was examined. B, Quantifications of TIMP-1 (healthy aorta [NA], 100 ± 8, versus AAA, 219 ± 9), TIMP-2 (NA, 100 ± 10, versus AAA, 14 ± 6), and TIMP-3 (NA, 100 ± 38, versus AAA, 385 ± 38) levels were performed with densitometry and are expressed as percent of values for healthy aorta. *Values of less than.05 were considered significant. Journal of Vascular Surgery 2002 35, 539-546DOI: (10.1067/mva.2002.121124) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 5 Matrix metalloproteinase 12 (MMP-12) protein expression and activity are increased in aortic aneurysms. A, MMP-12 expression was studied in healthy aorta (n = 3) and abdominal aortic aneurysms (AAAs; n = 5). Artery homogenates were resolved (25 μg protein/well) on 7.5% SDS-polyacrylamide electrophoresis gel, and MMP-12 immunoreactive forms were detected as described in Materials and Methods section. B, MMP-12 activity in artery homogenates was analyzed with casein zymography as described in Materials and Methods section. The 54-kd pro-MMP, 45-kd intermediate MMP-12 (iMMP-12), and 22-kd active forms of MMP-12 are identified. Standard used (Stds) was commercially available recombinant MMP-12. Journal of Vascular Surgery 2002 35, 539-546DOI: (10.1067/mva.2002.121124) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 6 Matrix metalloproteinase 2 (MMP-2) and MMP-9 gelatinolytic activities in healthy aorta and abdominal aortic aneurysms (AAAs). Homogenates of healthy aorta (n = 4) and AAAs (n = 5) were submitted (25 mg protein/well) to gelatin zymography as described in Materials and Methods section. Enzymatically active proteins were identified with clear bands against dark background of intact substrate. Positions of pro-MMP and active forms of MMP-2 and pro–MMP-9 are shown and indicate main gelatinolytic activities. Standards used (Stds; MMP-2 and MMP-9) were cultured medium from HT-1080 human fibrosarcoma cells. Journal of Vascular Surgery 2002 35, 539-546DOI: (10.1067/mva.2002.121124) Copyright © 2002 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions