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Ca overload and the action of calcium sensitive proteases, phospholipases and prostaglandin E2 in myocardial cell degradation

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Summary

The mechanism of Ca overload-induced myocardial cell injury under hypoxia was examined for the involvement of calcium-activated neutral proteases (CANP), calcium-dependent phospholipases (CDP) or prostaglandins with measuring45Ca entry, intake of biologically inert dye, nigrosin, into the cultured myocytes, as was useful for the quantification of sarcolemma permeability, and the release of creatine phosphokinase (CPK) to the culture medium. A Ca channel blocker, verapamil (1 and 10 μg/ml) or a Ca ionophore, A 23187 (0.5 to 4 μg/ml) dose-dependently decreased or increased both the Ca entry and nigrosin intake in accordance with the CPK release. Furthermore, the inhibitors against CANP, NCO-700 (2 and 20 μg/ml) that was demonstrated to permeate sarcolemma using14C-labelled reagent, against CDP, mepacrine (1 and 10 μg/ml) or against cyclooxygenase, indomethacin (1 and 10 μg/ml) caused no effect on the Ca entry, nigrosin intake nor CPK release under hypoxia. These results suggest that the Ca overdose into the myocardial cells potentiates their injury and it is not primarily related to the activation of CANP, CDP nor cyclooxygenase.

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References

  1. Acosta DM, Pucket DP, McMillan R (1978) Ischemic myocardial cell injury in cultured heart cells: Leakage of cytoplasmic enzyme from injured cells. In vitro 14:728–732

    PubMed  Google Scholar 

  2. Alonso DR, Scheid S, Post M, Killip T (1973) Pathophysiology of cardiogenic shock, quantification of myocardial necrosis, clinical, pathologic and electrocardiographic correlations. Circulation 48:588–596

    PubMed  Google Scholar 

  3. Barcos V, Rodeman HP, Dinarello CA, Goldberg AL (1983) Stimulation of muscle protein degradation and prostaglandin E2 release by leucocytic pyrogen (Interleukin-1). A mechanism for the increased degradation of muscle proteins. N Engl J Med 308:553–558

    PubMed  Google Scholar 

  4. Chien KR, Reeves JP, Buja M, Bonte F, Parkley RW, Willerson JT (1981) Phospholipid alterations in canine ischemic myocardium: Temporal topographic correlations with Tc-99m-PPi accumulation and in vitro sarcolemmal Ca2+ permeability. Circ Res 48:711–719

    PubMed  Google Scholar 

  5. Chizonite RA, Zak R (1980) Calcium-induced cell death: Susceptibility is age-dependent. Science 213:1508–1510

    Google Scholar 

  6. Chiariello M, Ambrosio G, Cappelli-Bigazzi M, Marone G, Conforelli G (1983) Phospholipase inhibition by quinacrine protects ischemic myocardium. Circulation 68(II, abstr):390

    Google Scholar 

  7. DeHaarn RL (1967) Regulation of spontaneous activity and growth of embryonic chicken heart cells in tissue culture. Develop Biol 16:216–249

    Article  PubMed  Google Scholar 

  8. Drummond GL, Duncan L (1980) The action of calcium ions on cardiac phosphorylase b kinase. J Biol Chem 241:3097–3103

    Google Scholar 

  9. Ebashi S (1980) Regulation of muscle contraction. A Croonian lecture. Proc Roy Soc London [Biol] 201:258–286

    Google Scholar 

  10. Fleckenstein A, Janke J, Doering HH, Leder O (1975) Key role of Ca in the production of non-coronarogenic myocardial necrosis. In: Fleckenstein A, Rona G (eds) Pathophysiology and morphology of myocardial alterations. University Park Press, Baltimore, p 21–42

    Google Scholar 

  11. Franson RC, Weir DL, Thakker J (1983) Solubilization and characterization of a neutral-active calcium-dependent phospholipase A2 from rabbit heart and lated chicken embryo myocytes. J Molec Cell Cardiol 15:189–196

    Article  Google Scholar 

  12. Huston RB, Krebs EG (1968) Activation of skeletal muscle phosphorylase kinase by Ca2+. II. Identification of the kinase activating factor as a proteolytic enzyme. Biochemistry 7(pt 2): 2116–2122

    Article  PubMed  Google Scholar 

  13. Kaltenbach JP, Kaltenbach MH, Lyons WB (1958) Nigrosin as a dye for differentiating live and dead ascites cells. Exp Cell Res 15:112–117

    Article  PubMed  Google Scholar 

  14. Kameyama T, Etlinger JD (1979) Calcium-dependent regulation of protein synthesis and degradation. Nature 279:344–346

    PubMed  Google Scholar 

  15. Kuroda M, Tanaka T, Masaki T (1981) Eu-actinin, a new structural protein of the z-lines of striated muscles. J Biochem 89:297–310

    PubMed  Google Scholar 

  16. Low MG, Weglicki WC (1982) Characterization of cytosolic Ca2+-dependent phosphatidylinositol specific phospholipase C in mammalian heart. Circulation 66(II, abstr): 110

    Google Scholar 

  17. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with Folin phenol reagent. J Biol Chem 193:265–275

    PubMed  Google Scholar 

  18. Mellgren RL (1980) Canine cardiac calcium-dependent proteases: Resolution of two forms with different requirement for calcium. FEBS Letters 109:129–133

    Article  PubMed  Google Scholar 

  19. Naylor WG, Grau A, Slade A (1976) A protective effect of verapamil on hypoxic heart muscle. Cardiovasc Res 10:650–662

    PubMed  Google Scholar 

  20. Naylor WG (1983) The role of calcium in myocardial ischemia and death. In: Stone PH, Antman EM (eds) Calcium channel blocking agents in the treatment of cardiovascular disorders. Futura Publ, New York, pp 81–105

    Google Scholar 

  21. Oliver IT (1955) A spectrophotometric method for the determination of creatine phosphokinase and myokinase. Biochem J 61:116–122

    PubMed  Google Scholar 

  22. Ozawa E, Hosoi K, Ebashi S (1967) Reversible stimulation of muscle phosphorylase b kinase by low concentration of calcium ions. J Biochem 61:531–533

    PubMed  Google Scholar 

  23. Page E, Polimeni PI (1977) Ultrastructural changes in the ischemic zone bordering experimental infarcts in rat ventricle. Am J Pathol 87:81–90

    PubMed  Google Scholar 

  24. Reddy MK, Etlinger JD, Rabinowitz M, Fischman DA, Zak R (1975) Removal of z-lines and alpha-actinin from isolated myofibrils by a calcium-activated neutral protease. J Biol Chem 250:4278–4284

    PubMed  Google Scholar 

  25. Renthrop, Blanke H, Koestering H, Karsch KR (1980) Intrakoronale Streptokinase-Applikation bei akutem Infarkt und instabiler Angina pectoris. Dtsch Med Wschr 105:221–228

    PubMed  Google Scholar 

  26. Rodeman HP, Waxman L, Goldberg AL (1982) The stimulation of protein degradation by Ca is mediated by prostaglandin E2 and does not require the calcium-activated neutral protease. J Biol Chem 257:8716–8723

    PubMed  Google Scholar 

  27. Shaikh NA, Downer E (1981) Time course of changes in porcine myocardial phospholipid levels during ischemia. Circ Res 49:316–325

    PubMed  Google Scholar 

  28. Shumway NE, Lower RR (1959) Topical cardiac hypothermia for extended period. Surg Forum 10:563–566

    Google Scholar 

  29. Smith AF (1979) Enzymes and routine diagnosis. In: Hearse DJ (ed) Enzymes in cardiology. DeLeires Publ, Chichester, pp 199–246

    Google Scholar 

  30. Toyo-oka T (1980) Increased activity of intramuscular proteases in the hyperthyroid state. FEBS Letters 117:122–124

    Article  PubMed  Google Scholar 

  31. Toyo-oka T (1981) Calcium ion-insensitive contraction of glycerinated porcine cardiac muscle fibers by MgITP. Circ Res 49:1350–1355

    PubMed  Google Scholar 

  32. Toyo-oka T (1982) Phosphorylation with cyclic adenosine 3′∶5′ monophosphate dependent protein kinase renders bovine cardiac troponin sensitive to the degradation by calcium-activated neutral protease. Biochem Biophys Res Commun 107:44–50

    Article  PubMed  Google Scholar 

  33. Toyo-oka T, Kamishiro T, Masaki T, Masaki M (1982) Reduction of experimentally produced acute myocardial infarction size by a new synthetic inhibitor, NCO-700, against calcium-activated neutral protense. Jpn Heart J 23:829–834

    PubMed  Google Scholar 

  34. Toyo-oka T, Masaki T (1979) Calcium-activated neutral protease from bovine ventricular muscle. J Molec Cell Cardiol 11:769–786

    Article  Google Scholar 

  35. Toyo-oka T, Nimmo LE, Ross J Jr (1981) Destruction of z-discs and eu-actinin in 0.1 mM Ca2+ and preservation by protease inhibitor, leupeptin. Circulation 64(II, abstr):61

    Google Scholar 

  36. Toyo-oka T, Okamoto J, Tanaka T, Masaki T (1980) Presence of myosin-like protein on liver cells, J Biochem 87:675–678

    PubMed  Google Scholar 

  37. Toyo-oka T, Ross J Jr (1981) Ca2+ sensitivity change and troponin loss in cardiac natural actomyosin after coronary occlusion. Am J Physiol 240(Heart and Circ Physiol):H704-H708

    PubMed  Google Scholar 

  38. Toyo-oka T, Shimizu T, Masaki T (1978) Inhibition of proteolytic activity of calcium-activated neutral protease by leupeptin and antipain. Biochem Biophys Res Commun 82:484–491

    Article  PubMed  Google Scholar 

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Author notes

  1. T. Toyo-oka M.D.

    Present address: Dept. of Cardiology, Jichi Medical School, 329-04, Minami-kawachi, Tochigi, (Japan)

Authors and Affiliations

  1. Department of Pharmacology, Institute of Basic Medical Sciences, School of Medicine, Tsukuba University, Ibaraki, (Japan)

    T. Toyo-oka M.D., K. Hara, N. Nakamura, M. Kitahara & T. Masaki

  2. Dept. of Cardiology, Jichi Medical School, 329-04, Minami-kawachi, Tochigi, (Japan)

    K. Hara, N. Nakamura, M. Kitahara & T. Masaki

Authors
  1. T. Toyo-oka M.D.
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  2. K. Hara
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  3. N. Nakamura
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  4. M. Kitahara
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  5. T. Masaki
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Additional information

A part of the present study was financially supported by the Ministry of Education, Culture and Science, the Ministry of Health and Welfare, and the Japanese Heart Foundation.

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Toyo-oka, T., Hara, K., Nakamura, N. et al. Ca overload and the action of calcium sensitive proteases, phospholipases and prostaglandin E2 in myocardial cell degradation. Basic Res Cardiol 80, 303–315 (1985). https://doi.org/10.1007/BF01907906

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  • DOI: https://doi.org/10.1007/BF01907906

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