title: Alcohol modulation of cardiac matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs favors collagen accumulation.
abstract: BACKGROUND
  Chronic alcohol consumption has been shown in human and animal studies to result in collagen accumulation, myocardial fibrosis, and heart failure. Cardiac fibroblasts produce collagen and regulate extracellular matrix (ECM) homeostasis through the synthesis and activity of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs), with the balance of MMPs/TIMPs determining the rate of collagen turnover. Dynamic changes of MMP and TIMP expression were reported in alcohol-induced hepatic fibrosis; however, the effect of alcohol on MMP/TIMP balance in the heart and cardiac fibroblasts is unknown. We hypothesized that alcohol exposure alters cardiac fibroblast MMP and TIMP expression to promote collagen accumulation in the heart.
  
  
  METHODS
  Cardiac fibroblasts isolated from adult rats were cultured in the presence of alcohol (12.5 to 200 mM) for 48 hours. MMP, TIMP, and collagen type I and III expression were assayed by Western blot analysis. Hydroxyproline (HPro) was used as a marker of collagen production. The in vivo cardiac effects of ethanol (EtOH) were determined using rats exposed to EtOH vapor for 2 weeks, resulting in blood alcohol levels of 150 to 200 mg/dl. Cardiac collagen volume fraction (CVF), as well as MMP, TIMP, and collagen expression, was assessed.
  
  
  RESULTS
  EtOH-exposed rats exhibited up-regulation of TIMP-1, TIMP-3 and TIMP-4 in the heart, with no significant increases in MMPs. Cardiac fibroblasts exhibited transformation to a profibrotic phenotype following exposure to alcohol. These changes were reflected by increased α-smooth muscle actin and collagen I and III expression, as well as increased collagen secretion. In vivo EtOH exposure also produced fibrosis, indicated by increased CVF and expression of collagens.
  
  
  CONCLUSIONS
  Alcohol exposure modulates cardiac fibroblast MMP/TIMP expression favoring a profile associated with collagen accumulation. Our data suggest that this disrupted MMP/TIMP profile may contribute to the development of myocardial fibrosis and cardiac dysfunction resulting from chronic alcohol abuse.
authors.affiliation: Department of Physiology , School of Medicine, Louisiana State University Health Science Center, New Orleans, Louisiana.
authors.forename: Elia C
authors.initials: EC
authors.lastname: El Hajj
authors.forename: Milad C
authors.initials: MC
authors.lastname: El Hajj
authors.forename: Tetyana G
authors.initials: TG
authors.lastname: Voloshenyuk
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Mouton
authors.forename: Elena
authors.initials: E
authors.lastname: Khoutorova
authors.forename: Patricia E
authors.initials: PE
authors.lastname: Molina
authors.forename: Nicholas W
authors.initials: NW
authors.lastname: Gilpin
authors.forename: Jason D
authors.initials: JD
authors.lastname: Gardner
mesh_terms: D000199:Actins / Q000378:metabolism; D000437:Alcoholism / Q000201:enzymology / Q000473:pathology; D000818:Animals; D015153:Blotting, Western; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D003094:Collagen / Q000378:metabolism*; D000431:Ethanol / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D005355:Fibrosis; D006321:Heart / Q000187:drug effects*; D006909:Hydroxyproline / Q000378:metabolism; D008297:Male; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D009206:Myocardium / Q000201:enzymology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019714:Tissue Inhibitor of Metalloproteinases / Q000378:metabolism*
chemical_list: D000199:Actins; D002492:Central Nervous System Depressants; D003094:Collagen; D000431:Ethanol; D006909:Hydroxyproline; D020782:Matrix Metalloproteinases; D019714:Tissue Inhibitor of Metalloproteinases; C498862:Acta2 protein, rat
keywords: Ethanol; Extracellular Matrix Remodeling; Fibrosis; Heart; Proteinase
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