Basic Research in Cardiology

, Volume 92, Issue 3, pp 147–158 | Cite as

Functional consequences of acute collagen degradation studied in crystalloid perfused rat hearts

  • K. Todaka
  • T. Jiang
  • J. T. Chapman
  • A. Gu
  • S. M. Zhu
  • E. Herzog
  • J. S. Hochman
  • S. F. Steinberg
  • D. Burkhoff
Original Contribution


Objectives: The impact of acute collagen disruption by the disulfide donor, 5,5′-dithio-2-nitrobenzoic acid (DTNB) on ventricular properties was tested in rat hearts.Methods: Collagen was degraded acutely in 13 isolated, isovlumically contracting rat hearts by perfusion with 1 mM DTNB added to Krebs-Henseleit solution for 1 hour followed by 2-hour perfusion with normal solution. Another 13 hearts were perfused with normal solution for 3 hours (Control).Results: Collagen content was 3.5±0.5% of ventricular dry weight in control group compared with 2.1±0.4% in DTNB group (decrease by 40%, p<0.01). Scanning electron micrographs revealed loss of the delicate collagen network surrounding muscle fibers in DTNB treated hearts. Developed pressure at a fixed volume decreased to 86±17% of the baseline value after 3-hour perfusion in the control group, whereas in DTNB treated hearts developed pressure fell to 68±13% (p<0.01). End-diastolic pressure was set at 5 mmHg at the beginning of the experiment and rose to 15±8 mmHg in control and 30±13 mmHg (p<0.01) in the treated hearts. Concomitantly, wet-to-dry weight ratio increased from 5.63±0.26 in control to 6.07±0.11 (p<0.05) in the DTNB treated hearts. A separate set of experiments on isolated myocytes excluded the possibility of a direct effect of DTNB on myocyte contractile function.Conclusions: These data suggested that with 40% collagen disruption by DTNB there is a significant increase in tissue edema that results in a decrease in chamber capacitance; in addition, there is a significant decrease in systolic performance which reflects the combined effect of edema and loss of collagen.

Key Words

Rat collagen developed pressure pressure-volume relationships extracellular matrix 


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  1. 1.
    Armstrong PW, Moe GW, Howard RJ, Grima EA, Cruz TF (1994) Structural remodelling in heart failure: gelatinase induction. Can J Cardiol 10: 214–220Google Scholar
  2. 2.
    Borg TK, Ranson WF, Moslehy FA, Caulfield JB (1981) Structural basis of ventricular stiffness. Lab Invest 44: 49–54Google Scholar
  3. 3.
    Burkhoff D, Weiss RG, Schulman SP, Kalil-Filho R, Wannenburg T, Gerstenblith G (1991) Influence of metabolic substrate on rat heart function and metabolism at different coronary flows. Am J Physiol 261 (Heart Circ Physiol 30): H741-H750Google Scholar
  4. 4.
    Caulfield JB, Borg TK (1979) The collagen network of the heart. Lab Invest 40: 364–372Google Scholar
  5. 5.
    Caulfield JB, Wolkowicz PE (1988) Inducible collagenolytic activity in isolated perfused rat heart. Am J Pathol 131: 199–205Google Scholar
  6. 6.
    Caulfield JB, Wolkowicz PE (1990) A mechanism for cardiac dilation. Heart Failure 6: 138–150Google Scholar
  7. 7.
    Cawston TE, Murphy G (1981) Mammalian collagenases. Methods in Enzymology 80: 711–722Google Scholar
  8. 8.
    Charney RH, Takahashi S, Zhao M, Sonnenblick E, Eng C (1992) Collagen loss in stunned myocardium. Circulation 85: 1483–1490Google Scholar
  9. 9.
    Dec GW, Palacios IF, Fallon JT, Aretz T, Mills J, Lee DC, Johnson RA (1985) Active myocarditis in the spectrum of acute dilated cardiomyopathies—clinical features, histologic correlates, and clinical outcome. N Engl J Med 312 (14): 885–890Google Scholar
  10. 10.
    Eng C, Zhao M, Factor SM, Sonnenblick EH (1993) Post-ischemic cardiac dilatation and remodeling: reperfusion injury of the interstitium. Europ Heart J 14 (Supplement A): 27–32Google Scholar
  11. 11.
    Factor SM, Robinson TF (1988) Comparative connective tissue structure-function relationships in biologic pumps. Lab Invest 58: 150–156Google Scholar
  12. 12.
    Kato S, Spinale FG, Tanaka R, Johnson W, Cooper GCI, Zile MR (1995) Inhibition of collagen cross-linking: effects on fibrillar collagen and ventricular diastolic function. Am J Physiol 269 (HCP 38): H863-H868Google Scholar
  13. 13.
    Kodama M, Hanawa H, Zhang S, Saeki M, Koyama S, Hosono H, Miyakita Y, Kotoh K, Inomata T, Izumi T, Shibata A (1993) FR506 therapy of experimental autoimmune myocarditis after onset of the disease. Am Heart J 126: 1385–1392Google Scholar
  14. 14.
    Komamura K, Shannon RP, Ihara T, Shen Y, Mirsky I, Bishop SP, Vatner SF (1993) Exnaustion of Frank-Starling mechanism in conscious dogs with heart failure. Am J Physiol 265 (Heart Circ Physiol 34): H1119-H1131Google Scholar
  15. 15.
    Kresh JY (1993) Intramyocardial mechanical states: vessel-interstitium-muscle inface. In: Interactive Phenomena in the Cardiac System, edited by S. Sideman and R. Beyar. New York: Plenum Press, p 113–123Google Scholar
  16. 16.
    Kuznetsov V, Pak E, Robinson RB, Steinberg SG (1995)β 2 receptor actions in neonatal and adult rat ventricular myocytes. Circ Res 76: 40–52Google Scholar
  17. 17.
    MacKenna DA, Omens JH, McCulloch AD, Covell JW (1994) Contribution of collagen matrix to passive left ventricular mechanics in isolated rat hearts. Am J Physiol 266 (Heart Circ Physiol 35): H1007-H1018Google Scholar
  18. 18.
    Matsubara BB, Henegar JR, Janicki JS (1992) Functional and morphological consequences of induced myocardial collagen damage. Circulation 86: I-171 (Abstract)Google Scholar
  19. 19.
    McDonald KS, Moss RL (1995) Osmotic compression of single cardiac myocytes eliminates the reduction in Ca2+ sensitivity of tension at short sarcomere length. Circ Res 77: 199–205Google Scholar
  20. 20.
    Robinson TF, Cohen-Gould L, Factor SM (1983) Skeletal framework of mammalian heart muscle: arrangement of inter-and pericellular connective tissue structures. Lab Invest 49: 482–498Google Scholar
  21. 21.
    Rubboli A, Sobotka PA, Euler DE (1994) Effect of acute edema on left ventricular function and coronary vascular resistance in the isolated rat heart. Am J Physiol 267 (Heart Circ Physiol 36): H1054-H1061Google Scholar
  22. 22.
    Sakai M, Danziger RS, Staddon JM, Lakatta EG, Hansford RG (1989) Decrease with senescence in the norepinephrine-induced phosphorylation of myofilament proteins in isolated rat cardiac myocytes. J Mol Cell Cardiol 21: 1327–1336Google Scholar
  23. 23.
    Spinale FG, Tomita M, Zellner JL, Cook JC, Crawford FA, Zile MR (1991) Colagen remodeling and changes in LV function during development and recovery from supraventricular tachycardia. Am J Physiol 261 (Heart Circ Physiol 30): H308-H318Google Scholar
  24. 24.
    Springman EB, Angleton EL, Birkedal-Hansen H, Van Wart HE (1990) Multiple mode of activation of latent human fibroblast collagenases: evidence for the role of a Cys73 active-site zinc complex in latency and a “cysteine switch” mechanism for activation. Proc Natl Acad Sci 87: 364–368Google Scholar
  25. 25.
    Stegemann H (1958) Mikrobestimmung von Hydroxyprolin mit Chloramin-T und p-Dimethylaminobenzaldehyd. Hoppe-seyleis 2 Physiol Chemie 311: 41–45Google Scholar
  26. 26.
    Takahashi S, Barry AC, Factor SM (1990) Collagen degradation in ischaemic rat hearts. Biochem J 265: 233–241Google Scholar
  27. 27.
    Weber KT (1989) Cardiac interstitium in health and disease: The fibrillar collagen network. J Am Coll Cardiol 13: 1637–1652Google Scholar
  28. 28.
    Weber KT, Janicki JS, Shroff SG, Pick R, Chen RM, Bashey RI (1988) Collagen remodeling of the pressure overloaded, hypertrophied nonhuman primate myocardium. Circ Res 62: 757–765Google Scholar
  29. 29.
    Weber KT, Pick R, Janicki JS, Gadodia G, Lakier JB (1988) Inadequate collagen tethers in dilated cardiomyopathy. Am Heart J 116: 1641–1646Google Scholar
  30. 30.
    Weber KT, Pick R, Silver MA, Moe GW, Janicki JS, Zucker IH, Armstrong PW (1990) Fibrillar collagen and remodeling of dilated canine left ventricle. Circulation 82: 1387–1404Google Scholar
  31. 31.
    Whittaker P, Boughner DR, Kloner RA (1991) Role of collagen in acute myocardial infarct expansion. Circulation 84: 2123–2134Google Scholar
  32. 32.
    Whittaker P, Boughner DR, Kloner RA, Przyklenk K (1991) Stunned myocardium and myocardial collagen damage: differential effects of single and repeated occlusions. Am Heart J 121: 434–441Google Scholar
  33. 33.
    Whittaker P, Kloner RA, Boughner DR, Pickering JG (1994) Quantitative assessment of myocardial collagen with picrosirius red staining and circularly polarized light. Basic Res Cardiol 89: 397–410Google Scholar
  34. 34.
    Woessner JF (1961) The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch Biochem Biophys 93: 440–447Google Scholar
  35. 35.
    Yoshikane H, Honda M, Goto Y, Morioka S, Ooshima A, Moriyama K (1992) Collagen in dilated cardiomyopathy — scanning electron microscopic and immunohistochemical observations. Jpn Circ J 56: 899–910Google Scholar
  36. 36.
    Zhao M, Zhang H, Robinson TF, Factor SM, Sonnenblick EH, Eng C (1987) Profound structural alterations of the extracellular collagen matrix in post-ischemic dysfunctional (“stunned”) but viable myocardium. J Am Coll Cardiol 10: 1322–1334Google Scholar
  37. 37.
    Zile MR, Izzi G, Gaasch WH (1991) Left ventricular diastolic dysfunction limits use of maximum systolic elastance as an index of contractile function. Circulation 83: 674–680Google Scholar

Copyright information

© Steinkopff Verlag 1997

Authors and Affiliations

  • K. Todaka
    • 1
  • T. Jiang
    • 2
  • J. T. Chapman
    • 1
  • A. Gu
    • 1
  • S. M. Zhu
    • 1
  • E. Herzog
    • 3
  • J. S. Hochman
    • 3
  • S. F. Steinberg
    • 1
  • D. Burkhoff
    • 1
  1. 1.Division of Circulatory PhysiologyNew YorkUSA
  2. 2.Department of PharmacologyCollege of Physicians & Surgeons of Columbia UniversityNew YorkUSA
  3. 3.Division of CardiologySt. Luke's/Roosevelt Hospital CenterNew YorkUSA

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