Skip to main content

Advertisement

SpringerLink
Log in

Reduction of myocardial infarct size by doxycycline: A role for plasmin inhibition

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Myocardial ischemia-reperfusion (I/R) is associated with the activation of matrix metalloproteinases (MMPs) and serine proteases. We hypothesized that activation of MMPs and the serine protease plasmin contribute to early cardiac myocyte death following I/R and that broad-spectrum protease inhibition with doxycycline (DOX) preserves myocyte viability. Rats treated daily with or without DOX beginning 48 h prior to experimentation were subjected to 30 min of coronary occlusion and 2 days of reperfusion. DOX pre-treatment reduced infarct size by 37%. DOX attenuated increases in MMP-9 and plasmin levels as determined by gelatin zymography and immunoblot, respectively. Neutrophil extravasation was unaltered by DOX as assessed by myeloperoxidase (MPO) activity. To examine the contribution of MMP-9 and plasmin to myocyte injury, cultures of neonatal rat ventricular myocytes (NRVMs) were treated for 48 h with 83 kDa MMP-9 or plasminogen in the presence or absence of DOX. MMP-9 treatment did not affect myocyte viability. Plasminogen treatment led to increased plasmin activity, resulting in loss of β1-integrin, NRVM detachment and apoptosis. DOX co-treatment inhibited plasmin activity and preserved NRVM attachment, whereas co-treatment with the broad-spectrum MMP inhibitor GM6001 had no effect. These results indicate that plasmin causes disruption of myocyte attachment and viability independently of MMP activation in vitro and that inhibition of plasmin by DOX may reduce I/R-induced myocyte death in vivo through the inhibition of plasmin. (Mol Cell Biochem 270: 1–11, 2005)

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Braunwald E: Myocardial reperfusion, limitation of infarct size, reduction of left ventricular dysfunction, and improved survival. Should the paradigm be expanded? Circulation 79: 441–444, 1989

    Google Scholar 

  2. Park JL, Lucchesi BR: Mechanisms of myocardial reperfusion injury. Ann Thorac Surg 68: 1905–1912, 1999

    Google Scholar 

  3. Kajstura J, Cheng W, Reiss K, Clark WA, Sonnenblick EH, Krajewski S, Reed JC, Olivetti G, Anversa P: Apoptotic and necrotic myocyte cell deaths are independent contributing variables of infarct size in rats. Lab Invest 74: 86–107, 1996

    CAS  PubMed  Google Scholar 

  4. Parsa CJ, Kim J, Riel RU, Pascal LS, Thompson RB, Petrofski JA, Matsumoto A, Stamler JS, Koch WJ: Cardioprotective effects of erythropoietin in the reperfused ischemic heart: a potential role for cardiac fibroblasts. J Biol Chem 279: 20655–20662, 2004

    Article  CAS  PubMed  Google Scholar 

  5. Stadler B, Phillips J, Toyoda Y, Federman M, Levitsky S, McCully JD: Adenosine-enhanced ischemic preconditioning modulates necrosis and apoptosis: Effects of stunning and ischemia-reperfusion. Ann Thorac Surg 72: 555–563; discussion 563–554, 2001

    Google Scholar 

  6. Wakiyama H, Cowan DB, Toyoda Y, Federman M, Levitsky S, McCully JD: Selective opening of mitochondrial ATP-sensitive potassium channels during surgically induced myocardial ischemia decreases necrosis and apoptosis. Eur J Cardiothorac Surg 21: 424–433, 2002

    Google Scholar 

  7. Sabri A, Alcott SG, Elouardighi H, Pak E, Derian C, Andrade-Gordon P, Kinnally K, Steinberg SF: Neutrophil cathepsin G promotes detachment-induced cardiomyocyte apoptosis via a protease-activated receptor-independent mechanism. J Biol Chem 278: 23944–23954, 2003

    Google Scholar 

  8. Ding B, Price RL, Goldsmith EC, Borg TK, Yan X, Douglas PS, Weinberg EO, Bartunek J, Thielen T, Didenko VV, Lorell BH: Left ventricular hypertrophy in ascending aortic stenosis mice: anoikis and the progression to early failure. Circulation 101: 2854–2862, 2000

    Google Scholar 

  9. Villarreal F, Omens J, Dillmann W, Risteli J, Nguyen J, Covell J: Early degradation and serum appearance of type I collagen fragments after myocardial infarction. J Mol Cell Cardiol 36: 597–601, 2004

    Google Scholar 

  10. Etoh T, Joffs C, Deschamps AM, Davis J, Dowdy K, Hendrick J, Baicu S, Mukherjee R, Manhaini M, Spinale FG: Myocardial and interstitial matrix metalloproteinase activity after acute myocardial infarction in pigs. Am J Physiol Heart Circ Physiol 281: H987–994, 2001

    Google Scholar 

  11. Nagase H, Woessner JF, Jr.: Matrix metalloproteinases. J Biol Chem 274: 21491–21494, 1999

    Google Scholar 

  12. Cheung PY, Sawicki G, Wozniak M, Wang W, Radomski MW, Schulz R: Matrix metalloproteinase-2 contributes to ischemia-reperfusion injury in the heart. Circulation 101: 1833–1839, 2000

    Google Scholar 

  13. Collen D: Ham-Wasserman lecture: Role of the plasminogen system in fibrin-homeostasis and tissue remodeling. Hematology (Am Soc Hematol Educ Program) 1–9, 2001

  14. Heymans S, Luttun A, Nuyens D, Theilmeier G, Creemers E, Moons L, Dyspersin GD, Cleutjens JP, Shipley M, Angellilo A, Levi M, Nube O, Baker A, Keshet E, Lupu F, Herbert JM, Smits JF, Shapiro SD, Baes M, Borgers M, Collen D, Daemen MJ, Carmeliet P: Inhibition of plasminogen activators or matrix metalloproteinases prevents cardiac rupture but impairs therapeutic angiogenesis and causes cardiac failure. Nat Med 5: 1135–1142, 1999

    Google Scholar 

  15. Knoepfler PS, Bloor CM, Carroll SM: Urokinase plasminogen activator activity is increased in the myocardium during coronary artery occlusion. J Mol Cell Cardiol 27: 1317–1324, 1995

    Google Scholar 

  16. Gurevitch J, Barak J, Hochhauser E, Paz Y, Yakirevich V: Aprotinin improves myocardial recovery after ischemia and reperfusion. Effects of the drug on isolated rat hearts. J Thorac Cardiovasc Surg 108: 109–118, 1994

    Google Scholar 

  17. McCarthy RJ, Tuman KJ, O’Connor C, Ivankovich AD: Aprotinin pretreatment diminishes postischemic myocardial contractile dysfunction in dogs. Anesth Analg 89: 1096–1100, 1999

    Google Scholar 

  18. Golub LM, Lee HM, Ryan ME, Giannobile WV, Payne J, Sorsa T: Tetracyclines inhibit connective tissue breakdown by multiple non-antimicrobial mechanisms. Adv Dent Res 12: 12–26, 1998

    Google Scholar 

  19. Imamura T, Matsushita K, Travis J, Potempa J: Inhibition of trypsin-like cysteine proteinases (gingipains) from Porphyromonas gingivalis by tetracycline and its analogues. Antimicrob Agents Chemother 45: 2871–2876, 2001

    Google Scholar 

  20. Sorsa T, Lindy O, Konttinen YT, Suomalainen K, Ingman T, Saari H, Halinen S, Lee HM, Golub LM, Hall J, et al.: Doxycycline in the protection of serum alpha-1-antitrypsin from human neutrophil collagenase and gelatinase. Antimicrob Agents Chemother 37: 592–594, 1993

    Google Scholar 

  21. Grenier D, Plamondon P, Sorsa T, Lee HM, McNamara T, Ramamurthy NS, Golub LM, Teronen O, Mayrand D: Inhibition of proteolytic, serpinolytic, and progelatinase-b activation activities of periodontopathogens by doxycycline and the non-antimicrobial chemically modified tetracycline derivatives. J Periodontol 73: 79–85, 2002

    Google Scholar 

  22. Curci JA, Petrinec D, Liao S, Golub LM, Thompson RW: Pharmacologic suppression of experimental abdominal aortic aneurysms: Acomparison of doxycycline and four chemically modified tetracyclines. J Vasc Surg 28: 1082–1093, 1998

    Google Scholar 

  23. Lamparter S, Slight SH, Weber KT: Doxycycline and tissue repair in rats. J Lab Clin Med 139: 295–302, 2002

    Google Scholar 

  24. Tiefenbacher CP, Kapitza J, Dietz V, Lee CH, Niroomand F: Reduction of myocardial infarct size by fluvastatin. Am J Physiol Heart Circ Physiol 285: H59–64, 2003

    Google Scholar 

  25. Hawkes SP, Li H, Taniguchi GT: Zymography and reverse zymography for detecting MMPs and TIMPs. In: I. M. Clark, (ed.). Matrix Metalloproteinase Protocols, Humana Press, Inc., Totowa, NJ, 2001, pp. 399–410

    Google Scholar 

  26. Romanic AM, Harrison SM, Bao W, Burns-Kurtis CL, Pickering S, Gu J, Grau E, Mao J, Sathe GM, Ohlstein EH, Yue TL: Myocardial protection from ischemia/reperfusion injury by targeted deletion of matrix metalloproteinase-9. Cardiovasc Res 54: 549–558, 2002

    Google Scholar 

  27. Pozzi A, Moberg PE, Miles LA, Wagner S, Soloway P, Gardner HA: Elevated matrix metalloprotease and angiostatin levels in integrin alpha 1 knockout mice cause reduced tumor vascularization. Proc Natl Acad Sci U.S.A. 97: 2202–2207, 2000

    Google Scholar 

  28. Pham CG, Harpf AE, Keller RS, Vu HT, Shai SY, Loftus JC, Ross RS: Striated muscle-specific beta(1D)-integrin and FAK are involved in cardiac myocyte hypertrophic response pathway. Am J Physiol Heart Circ Physiol 279: H2916–2926, 2000

    Google Scholar 

  29. Iwaki K, Sukhatme VP, Shubeita HE, Chien KR: Alpha- and beta-adrenergic stimulation induces distinct patterns of immediate early gene expression in neonatal rat myocardial cells. fos/jun expression is associated with sarcomere assembly; Egr-1 induction is primarily an alpha 1-mediated response. J Biol Chem 265: 13809–13817, 1990

    Google Scholar 

  30. Meilhac O, Ho-Tin-Noe B, Houard X, Philippe M, Michel JB, Angles-Cano E: Pericellular plasmin induces smooth muscle cell anoikis. Faseb J 17: 1301–1303, 2003

    Google Scholar 

  31. Lindsey M, Wedin K, Brown MD, Keller C, Evans AJ, Smolen J, Burns AR, Rossen RD, Michael L, Entman M: Matrix-dependent mechanism of neutrophil-mediated release and activation of matrix metalloproteinase 9 in myocardial ischemia/reperfusion. Circulation 103: 2181–2187, 2001

    Google Scholar 

  32. Griswold DE, Hillegass LM, Hill DE, Egan JW, Smith EF, 3rd: Method for quantification of myocardial infarction and inflammatory cell infiltration in rat cardiac tissue. J Pharmacol Methods 20: 225–235, 1988

    Google Scholar 

  33. Kaden JJ, Dempfle CE, Sueselbeck T, Brueckmann M, Poerner TC, Haghi D, Haase KK, Borggrefe M: Time-dependent changes in the plasma concentration of matrix metalloproteinase 9 after acute myocardial infarction. Cardiology 99: 140–144, 2003

    Google Scholar 

  34. Ross RS, Borg TK: Integrins and the myocardium. Circ Res 88: 1112–1119, 2001

    Google Scholar 

  35. Smith EF, 3rd, Egan JW, Bugelski PJ, Hillegass LM, Hill DE, Griswold DE: Temporal relation between neutrophil accumulation and myocardial reperfusion injury. Am J Physiol 255: H1060–1068, 1988

    Google Scholar 

  36. Kaito K, Urayama H, Watanabe G: Doxycycline treatment in a model of early abdominal aortic aneurysm. Surg Today 33: 426–433, 2003

    Google Scholar 

  37. Heidkamp MC, Bayer AL, Kalina JA, Eble DM, Samarel AM: GFP-FRNK disrupts focal adhesions and induces anoikis in neonatal rat ventricular myocytes. Circ Res 90: 1282–1289, 2002

    Google Scholar 

  38. Owens MR, Cimino CD: Biosynthesis of plasminogen by the perfused rat liver. J Lab Clin Med 105: 368–373, 1985

    Google Scholar 

  39. Creemers E, Cleutjens J, Smits J, Heymans S, Moons L, Collen D, Daemen M, Carmeliet P: Disruption of the plasminogen gene in mice abolishes wound healing after myocardial infarction. Am J Pathol 156: 1865–1873, 2000

    Google Scholar 

  40. Rossignol P, Ho-Tin-Noe B, Vranckx R, Bouton MC, Meilhac O, Lijnen HR, Guillin MC, Michel JB, Angles-Cano E: Protease-nexin-1 inhibits plasminogen activation-induced apoptosis of adherent cells. J Biol Chem 279: 10346–10356, 2003

    Google Scholar 

  41. Reinartz J, Schafer B, Batrla R, Klein CE, Kramer MD: Plasmin abrogates alpha v beta 5-mediated adhesion of a human keratinocyte cell line (HaCaT) to vitronectin. Exp Cell Res 220: 274–282, 1995

    Google Scholar 

  42. Gately S, Twardowski P, Stack MS, Cundiff DL, Grella D, Castellino FJ, Enghild J, Kwaan HC, Lee F, Kramer RA, Volpert O, Bouck N, Soff GA: The mechanism of cancer-mediated conversion of plasminogen to the angiogenesis inhibitor angiostatin. Proc Natl Acad Sci USA 94: 10868–10872, 1997

    Google Scholar 

  43. Strickland DK, Kounnas MZ, Argraves WS: LDL receptor-related protein: a multiligand receptor for lipoprotein and proteinase catabolism. Faseb J 9: 890–898, 1995

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Medicine, University of California, San Diego, CA, USA

    Michael O. Griffin, Miki Jinno & Francisco J. Villarreal MD PhD, UCSD

  2. Department of Cell Biology, The Scripps Research Institute, La Jolla, CA, USA

    Lindsey A. Miles

  3. UCSD Medical Center, M/C 8412, 200West Arbor Drive, San Diego, CA, 92103-8412, U.S.A.

    Francisco J. Villarreal MD PhD, UCSD

Authors
  1. Michael O. Griffin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Miki Jinno
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lindsey A. Miles
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Francisco J. Villarreal MD PhD, UCSD
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Francisco J. Villarreal MD PhD, UCSD.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Griffin, M.O., Jinno, M., Miles, L.A. et al. Reduction of myocardial infarct size by doxycycline: A role for plasmin inhibition. Mol Cell Biochem 270, 1–11 (2005). https://doi.org/10.1007/s11010-005-2540-3

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-005-2540-3

Key words

Search

Navigation