Genetic deficiency of plasminogen activator inhibitor-1 promotes cardiac fibrosis in aged mice

Involvement of constitutive transforming growth factor-β signaling and endothelial-to-mesenchymal transition

Asish K Ghosh, William S. Bradham, Linda A. Gleaves, Bart De Taeye, Sheila B. Murphy, Joseph W. Covington, Douglas E Vaughan

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

Background: Elevated levels of plasminogen activator inhibitor-1 (PAI-1), a potent inhibitor of urokinase plasminogen activator and tissue plasminogen activator, are implicated in the pathogenesis of tissue fibrosis. Paradoxically, lack of PAI-1 in the heart is associated with the development of cardiac fibrosis in aged mice. However, the molecular basis of cardiac fibrosis in aged PAI-1-deficient mice is unknown. Here, we investigated the molecular and cellular bases of myocardial fibrosis. Methods and Results: Histological evaluation of myocardial tissues derived from aged PAI-1-deficient mice revealed myocardial fibrosis resulting from excessive accumulation of collagen. Immunohistochemical characterization revealed that the levels of matrix metalloproteinase-2, matrix metalloproteinase-9, and transforming growth factor-β1/2 and the number of Mac3-positive and fibroblast specific protein-1-positive cells were significantly elevated in aged PAI-1-deficient myocardial tissues compared with controls. Zymographic analysis revealed that matrix metalloproteinase-2 enzymatic activity was elevated in PAI-1-deficient mouse cardiac endothelial cells. Real-time quantitative polymerase chain reaction analyses of RNA from myocardial tissues revealed the upregulation of profibrotic markers in aged PAI-1-deficient mice. The numbers of phosphorylated Smad2-, phosphorylated Smad3-, and phosphorylated ERK1/2 MAPK-, but not pAkt/PKB-, positive cells were significantly increased in PAI-1-deficient myocardial tissues. Western blot and immunocytochemical analysis revealed that PAI-1-deficient mouse cardiac endothelial cells were more susceptible to endothelial-to-mesenchymal transition in response to transforming growth factor-β2. Conclusions: These results indicate that spontaneous activation of both Smad and non-Smad transforming growth factor-β signaling may contribute to profibrotic responses in aged PAI-1-deficient mice hearts and establish a possible link between endothelial-to-mesenchymal transition and cardiac fibrosis in PAI-1-deficient mice.

Original languageEnglish (US)
Pages (from-to)1200-1209
Number of pages10
JournalCirculation
Volume122
Issue number12
DOIs
StatePublished - Sep 21 2010

Fingerprint

Plasminogen Activator Inhibitor 1
Transforming Growth Factors
Fibrosis
Matrix Metalloproteinase 2
Plasminogen Activator Inhibitor-1 Deficiency
Endothelial Cells
Plasminogen Inactivators
Matrix Metalloproteinase 9
Tissue Plasminogen Activator
Real-Time Polymerase Chain Reaction
Up-Regulation
Collagen
Western Blotting
RNA

Keywords

  • fibrosis
  • heart
  • inflammation
  • MMP
  • PAI-1
  • Smad
  • TGF-β

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

@article{a244cc5bf4534ddf9f8cdbbc44f07b1b,
title = "Genetic deficiency of plasminogen activator inhibitor-1 promotes cardiac fibrosis in aged mice: Involvement of constitutive transforming growth factor-β signaling and endothelial-to-mesenchymal transition",
abstract = "Background: Elevated levels of plasminogen activator inhibitor-1 (PAI-1), a potent inhibitor of urokinase plasminogen activator and tissue plasminogen activator, are implicated in the pathogenesis of tissue fibrosis. Paradoxically, lack of PAI-1 in the heart is associated with the development of cardiac fibrosis in aged mice. However, the molecular basis of cardiac fibrosis in aged PAI-1-deficient mice is unknown. Here, we investigated the molecular and cellular bases of myocardial fibrosis. Methods and Results: Histological evaluation of myocardial tissues derived from aged PAI-1-deficient mice revealed myocardial fibrosis resulting from excessive accumulation of collagen. Immunohistochemical characterization revealed that the levels of matrix metalloproteinase-2, matrix metalloproteinase-9, and transforming growth factor-β1/2 and the number of Mac3-positive and fibroblast specific protein-1-positive cells were significantly elevated in aged PAI-1-deficient myocardial tissues compared with controls. Zymographic analysis revealed that matrix metalloproteinase-2 enzymatic activity was elevated in PAI-1-deficient mouse cardiac endothelial cells. Real-time quantitative polymerase chain reaction analyses of RNA from myocardial tissues revealed the upregulation of profibrotic markers in aged PAI-1-deficient mice. The numbers of phosphorylated Smad2-, phosphorylated Smad3-, and phosphorylated ERK1/2 MAPK-, but not pAkt/PKB-, positive cells were significantly increased in PAI-1-deficient myocardial tissues. Western blot and immunocytochemical analysis revealed that PAI-1-deficient mouse cardiac endothelial cells were more susceptible to endothelial-to-mesenchymal transition in response to transforming growth factor-β2. Conclusions: These results indicate that spontaneous activation of both Smad and non-Smad transforming growth factor-β signaling may contribute to profibrotic responses in aged PAI-1-deficient mice hearts and establish a possible link between endothelial-to-mesenchymal transition and cardiac fibrosis in PAI-1-deficient mice.",
keywords = "fibrosis, heart, inflammation, MMP, PAI-1, Smad, TGF-β",
author = "Ghosh, {Asish K} and Bradham, {William S.} and Gleaves, {Linda A.} and {De Taeye}, Bart and Murphy, {Sheila B.} and Covington, {Joseph W.} and Vaughan, {Douglas E}",
year = "2010",
month = "9",
day = "21",
doi = "10.1161/CIRCULATIONAHA.110.955245",
language = "English (US)",
volume = "122",
pages = "1200--1209",
journal = "Circulation",
issn = "0009-7322",
publisher = "Lippincott Williams and Wilkins",
number = "12",

}

Genetic deficiency of plasminogen activator inhibitor-1 promotes cardiac fibrosis in aged mice : Involvement of constitutive transforming growth factor-β signaling and endothelial-to-mesenchymal transition. / Ghosh, Asish K; Bradham, William S.; Gleaves, Linda A.; De Taeye, Bart; Murphy, Sheila B.; Covington, Joseph W.; Vaughan, Douglas E.

In: Circulation, Vol. 122, No. 12, 21.09.2010, p. 1200-1209.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Genetic deficiency of plasminogen activator inhibitor-1 promotes cardiac fibrosis in aged mice

T2 - Involvement of constitutive transforming growth factor-β signaling and endothelial-to-mesenchymal transition

AU - Ghosh, Asish K

AU - Bradham, William S.

AU - Gleaves, Linda A.

AU - De Taeye, Bart

AU - Murphy, Sheila B.

AU - Covington, Joseph W.

AU - Vaughan, Douglas E

PY - 2010/9/21

Y1 - 2010/9/21

N2 - Background: Elevated levels of plasminogen activator inhibitor-1 (PAI-1), a potent inhibitor of urokinase plasminogen activator and tissue plasminogen activator, are implicated in the pathogenesis of tissue fibrosis. Paradoxically, lack of PAI-1 in the heart is associated with the development of cardiac fibrosis in aged mice. However, the molecular basis of cardiac fibrosis in aged PAI-1-deficient mice is unknown. Here, we investigated the molecular and cellular bases of myocardial fibrosis. Methods and Results: Histological evaluation of myocardial tissues derived from aged PAI-1-deficient mice revealed myocardial fibrosis resulting from excessive accumulation of collagen. Immunohistochemical characterization revealed that the levels of matrix metalloproteinase-2, matrix metalloproteinase-9, and transforming growth factor-β1/2 and the number of Mac3-positive and fibroblast specific protein-1-positive cells were significantly elevated in aged PAI-1-deficient myocardial tissues compared with controls. Zymographic analysis revealed that matrix metalloproteinase-2 enzymatic activity was elevated in PAI-1-deficient mouse cardiac endothelial cells. Real-time quantitative polymerase chain reaction analyses of RNA from myocardial tissues revealed the upregulation of profibrotic markers in aged PAI-1-deficient mice. The numbers of phosphorylated Smad2-, phosphorylated Smad3-, and phosphorylated ERK1/2 MAPK-, but not pAkt/PKB-, positive cells were significantly increased in PAI-1-deficient myocardial tissues. Western blot and immunocytochemical analysis revealed that PAI-1-deficient mouse cardiac endothelial cells were more susceptible to endothelial-to-mesenchymal transition in response to transforming growth factor-β2. Conclusions: These results indicate that spontaneous activation of both Smad and non-Smad transforming growth factor-β signaling may contribute to profibrotic responses in aged PAI-1-deficient mice hearts and establish a possible link between endothelial-to-mesenchymal transition and cardiac fibrosis in PAI-1-deficient mice.

AB - Background: Elevated levels of plasminogen activator inhibitor-1 (PAI-1), a potent inhibitor of urokinase plasminogen activator and tissue plasminogen activator, are implicated in the pathogenesis of tissue fibrosis. Paradoxically, lack of PAI-1 in the heart is associated with the development of cardiac fibrosis in aged mice. However, the molecular basis of cardiac fibrosis in aged PAI-1-deficient mice is unknown. Here, we investigated the molecular and cellular bases of myocardial fibrosis. Methods and Results: Histological evaluation of myocardial tissues derived from aged PAI-1-deficient mice revealed myocardial fibrosis resulting from excessive accumulation of collagen. Immunohistochemical characterization revealed that the levels of matrix metalloproteinase-2, matrix metalloproteinase-9, and transforming growth factor-β1/2 and the number of Mac3-positive and fibroblast specific protein-1-positive cells were significantly elevated in aged PAI-1-deficient myocardial tissues compared with controls. Zymographic analysis revealed that matrix metalloproteinase-2 enzymatic activity was elevated in PAI-1-deficient mouse cardiac endothelial cells. Real-time quantitative polymerase chain reaction analyses of RNA from myocardial tissues revealed the upregulation of profibrotic markers in aged PAI-1-deficient mice. The numbers of phosphorylated Smad2-, phosphorylated Smad3-, and phosphorylated ERK1/2 MAPK-, but not pAkt/PKB-, positive cells were significantly increased in PAI-1-deficient myocardial tissues. Western blot and immunocytochemical analysis revealed that PAI-1-deficient mouse cardiac endothelial cells were more susceptible to endothelial-to-mesenchymal transition in response to transforming growth factor-β2. Conclusions: These results indicate that spontaneous activation of both Smad and non-Smad transforming growth factor-β signaling may contribute to profibrotic responses in aged PAI-1-deficient mice hearts and establish a possible link between endothelial-to-mesenchymal transition and cardiac fibrosis in PAI-1-deficient mice.

KW - fibrosis

KW - heart

KW - inflammation

KW - MMP

KW - PAI-1

KW - Smad

KW - TGF-β

UR - http://www.scopus.com/inward/record.url?scp=77957278506&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77957278506&partnerID=8YFLogxK

U2 - 10.1161/CIRCULATIONAHA.110.955245

DO - 10.1161/CIRCULATIONAHA.110.955245

M3 - Article

VL - 122

SP - 1200

EP - 1209

JO - Circulation

JF - Circulation

SN - 0009-7322

IS - 12

ER -