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Biochemistry (Moscow)

, Volume 82, Issue 2, pp 168–175 | Cite as

Chronic alcohol intoxication is not accompanied by an increase in calpain proteolytic activity in cardiac muscle of rats

  • Yu. V. Gritsyna
  • N. N. Salmov
  • A. G. Bobylev
  • I. S. Fadeeva
  • N. I. Fesenko
  • D. G. Sadikova
  • N. I. Kukushkin
  • Z. A. Podlubnaya
  • I. M. VikhlyantsevEmail author
Article

Abstract

Enzymatic activity of Ca2+-dependent calpain proteases as well as the content and gene expression of μ-calpain (activated by micromolar calcium ion concentrations), calpastatin (inhibitor of calpains), and titin (substrate for calpains) were investigated in cardiac muscles of rats subjected to chronic alcoholization for 3 and 6 months. There was no increase in the “heart weight/body weight” parameter indicating development of heart hypertrophy in the alcoholized rats, while a decreasing trend was observed for this parameter in the rats after 6-month modeling of alcoholic cardiomyopathy, which indicated development of atrophic changes in the myocardium. Fluorometric measurements conducted using the Calpain Activity Assay Kit did not reveal any changes in total calpain activity in protein extracts of cardiac muscles of the rats alcoholized for 3 and 6 months. Western blot analysis did not show reliable changes in the contents of μ-calpain and calpastatin, and SDS-PAGE did not reveal any decrease in the titin content in the myocardium of rats after the chronic alcohol intoxication. Autolysis of μ-calpain was also not verified, which could indicate that proteolytic activity of this enzyme in myocardium of chronically alcoholized rats is not enhanced. Using Pro-Q Diamond staining, changes in phosphorylation level of titin were not detected in cardiac muscle of rats after chronic alcoholization during three and six months. A decrease in μ-calpain and calpastatin mRNA content (~1.3-fold, p ≤ 0.01 and ~1.9-fold, p ≤ 0.01, respectively) in the myocardium of rats alcoholized for 3 months and decrease in calpastatin mRNA (~1.4-fold, p ≤ 0.01) in animals alcoholized for 6 months was demonstrated using real-time PCR. These results indicate negative effect of chronic alcohol intoxication on expression of the abovementioned genes.

Keywords

cardiac muscle μ-calpain calpastatin titin chronically alcoholized rats 

Abbreviations

MHC

myosin heavy chains

T1

intact titin molecule located between the M-band and Z-disk of a sarcomere

T2

proteolytic fragment of the intact T1 linked to myosin filaments in A-disk of a sarcomere

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Copyright information

© Pleiades Publishing, Ltd. 2017

Authors and Affiliations

  • Yu. V. Gritsyna
    • 1
  • N. N. Salmov
    • 1
  • A. G. Bobylev
    • 1
  • I. S. Fadeeva
    • 1
  • N. I. Fesenko
    • 1
  • D. G. Sadikova
    • 2
  • N. I. Kukushkin
    • 2
  • Z. A. Podlubnaya
    • 1
    • 3
  • I. M. Vikhlyantsev
    • 1
    • 3
    Email author
  1. 1.Institute of Theoretical and Experimental BiophysicsRussian Academy of SciencesPushchino, Moscow RegionRussia
  2. 2.Institute of Cell BiophysicsPushchino, Moscow RegionRussia
  3. 3.Pushchino State Institute of Natural SciencesPushchino, Moscow RegionRussia

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