Chronic Alcohol Intoxication Is Not Accompanied by an Increase in Calpain Proteolytic Activity in Cardiac Muscle of Rats
| Autor: | Ivan M. Vikhlyantsev, A. G. Bobylev, Zoya A. Podlubnaya, N. N. Salmov, I S Fadeeva, Yu. V. Gritsyna, N. I. Fesenko, D. G. Sadikova, N. I. Kukushkin |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Male medicine.medical_specialty chemistry.chemical_element Muscle Proteins Apoptosis Alcoholic cardiomyopathy Calcium Biochemistry 03 medical and health sciences Western blot Internal medicine Gene expression medicine Animals Rats Wistar Calpastatin biology medicine.diagnostic_test Calpain Cardiomyopathy Alcoholic Myocardium Cardiac muscle General Medicine medicine.disease Molecular biology Rats 030104 developmental biology Endocrinology medicine.anatomical_structure chemistry Chronic Disease Proteolysis biology.protein Titin Alcoholic Intoxication |
| Zdroj: | Biochemistry. Biokhimiia. 82(2) |
| ISSN: | 1608-3040 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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