Histologic Features of Rat Gastrocnemius Muscle Under Conditions of Experimental Ischemia of Different Duration Complicated by Chronic Alcoholization





gastrocnemius muscle, ischemia, chronic alcoholization, pathomorphological changes, muscle fibers, endomysium, sarcolema


The purpose of the study was the histological changes analysis in the gastrocnemius muscle under the conditions of experimental ischemia of different duration complicated by chronic alcoholization. The experiment was conducted on inbred Wistar line rats with 140 – 160 g body weight. The animals were divided into four groups: control; animals with ischemia (1, 2 and 3 hours ischemia duration); alcoholized animals and alcoholized animals with experimentally induced 1, 2, 3 hours ischemia. Chronic alcohol intoxication of the animals was achieved by intragastric delivery of 40 % ethyl alcohol for 30 days. Unilateral vascular ischemia was induced by tourniquet ligating of the main arteries. Histologic processing of the tissue was carried out according to standard histological methods, stained with Van Gison method. Morphometric indices (thickness of muscle fibers and the size of interfibrillary space) were measured using VideoTest Morphology 5,0 software. It was found that the level of expression of morphological changes directly depends on the duration of ischemia. With its increase, destructive changes of the gastrocnemius muscle become more evident: invaginations on the sarcolmes of fibers are more clearly visible, myofibrils are disorganized, the amount of collagen increases and the scar formation starts. After three-hour ishemia the cytological signs of necrosis are observed, but in many areas the typical cross-striped and the normal structure of muscle fibers was preserved. Under the conditions of chronic alcoholization and ischemia of different duration, the structure and course of partially atrophied after alcoholic myopathy fibers are broken, degenerative changes in the integrity of sarcolemes are observed, some fibers decrease the volume of myofibrils, there endomysium swells and its necrosis starts, myofibrils dissociate. The presence of neutrophils between myofibrils and diffuse hemorrhage has been observed, nevertheless, in some sites a typical cross-striped structure was preserved. The obtained results show the negative synergy of myotoxic effects of ethyl alcohol and the destructive effects of vascular ischemia on fibers of rat m. gastrocnemius.


1. Murdock, M. Compartment syndrome: a review of the literature. Clin. Podiatr. Med. Surg; 2012, 29 (2), p 301–310.
2. Заводський, Д.; Ноздренко, Д.; Хома О.; Сорока, В. Зміна швидкісно-силових показників скорочення гомілкового м’язу щура за умов штучно викликаної васкулярної ішемії. Вісник Київського університету імені Тараса Шевченка. Серія біологічна; 2013, 63, с 5–7. https://doi.org/10.31558/1815-3070.2018.36.18
3. Martin, F. C.; Slavin, G.; Levi, A. J.; Peters T. J. Investigation of the organelle pathology of skeletal muscle in chronic alcoholism. J Clin. Pathol; 1984, 37, рр 448–454. https://doi.org/10.1136/jcp.37.4.448
4. Erkut, B.; Özyazıcıoğlu, A.; Karapolat, B. S.; Koçoğulları, C..U.; Keles, S.; Ateş, A.; Gundogdu, C.; Kocak, H. Effects of ascorbic аcid, alpha-tocopherol and allopurinol on ischemia-reperfusion injury in rabbit skeletal muscle: an experimental stud. Drug Target Insights; 2007; 2, p 249–58. https://doi.org/10.4137/dti.s303
5. Rácz, I. B.; Illyés, G.; Sarkadi, L.; Hamar J. The functional and morphological damage of ischemic reperfused skeletal muscle. Eur Surg Res; 1997, 29 (4), pp 254–63. https://doi.org/10.1159/000129531
6. Duarte, J. A.; Gloser, S.; Remiao, F.; Carvalho, F.; Bastos, M. L.; Soares, J. M.; Appell, H. J. Administration of tourniquet: I. Are edema and oxidative stress related to eachother and to the duration of ischemia in reperfused skeletal muscle? Arch Orthop Trauma Surg; 116, 1997, pp 97–100. https://doi.org/10.1007/bf00434110
7. Noveli, G. P.; Adembri, C.; Gandini, E.; Orlandini, S. Z.; Papucci, L.; Formigli, L.; Manneschi, L. I.; Quattrone, A.; Patresi, C.; Capaccioli, S. Vitamin E protects human skeletal muscle from damage during surgical ischemia-reperfusion. Am J Surg; 1997; 173, pp 206–209. https://doi.org/10.1016/s0002-9610(97)89593-1
8. Халилов, М. Х.; Закихорджаев, Ш. Я. К характеристике некоторых патохимических сдвигов в крови, тканях печени и головного мозга при экспериментальной алкогольной интоксикации; Вопросы клиники алкоголизма: сб. науч. тр., Ташкент, 1983; c 38–41.
9. Меркулов, Г. А. Курс паталогической техники, 3 изд, исп. и доп.; Медгиз: Ленинград, 1956; с 124–126.
10. Fernandez-Sola, J.; Preedy, V. R.; Lang, C. H.; Gonzalez-Reimers, E.; Arno, M.; Lin, J. C. I.; Wiseman, H.; Zhou, S.; Emery, P. W.; Nakahara, T. Molecular and cellular events in alcohol-induced muscle disease. Alcohol Clin Exp Res; 2007, 31 (12), pp 1953–1962. https://doi.org/10.1111/j.1530-0277.2007.00530.x
11. Дереча, Л. М. Алкоголь та його дія на організм: огляд літератури. Вісник Харківського національного університету ім. В. Н. Каразіна. Серія: біологія; 2007, 6 (788), c 7–16.
12. Hunter, R. J.; Neagoe, C.; Järveläinen, H. A.; Martin, C. R.; Lindros, K. O.; Linke, W. A.; Preedy, V. R. Alcohol affects the skeletal muscle proteins, titin and nebulin in male and female rats. J. Nutr; 2003, 133 (4), pp 1154 –1157.
13. Preedy, V. R.; Adachi, J.; Peters, T. J.; Worrall, S.; Parkkila, S.; Niemela, O.; Asano, M.; Ueno, Y.; Takeda, K.; Yamauchi, M.; et al. Recent advances in the pathology of alcoholic myopathy. Alcohol Clin Exp Res; 2001, 25 (5), pp 54S–59S. https://doi.org/10.1111/j.1530-0277.2001.tb02374.x
14. Preedy, V. R.; Adachi, J.; Veno, G.; Ahmed, S.; Mantle, D.; Mullatti, N.; Rajendram, R.; Peters, T. J. Alcoholic skeletal muscle myopathy: definitions, features, contribution of neuropathy, impact and diagnosis. Eur. J. Neurol; 2001, 8 (6), pp 677–687. https://doi.org/10.1046/j.1468-1331.2001.00303.x
15. Preedy, V. R.; Paice, A.; Mantle, D.; Dhillon, A. S.; Palmer, T. N.; Peters, T. J.; Alcoholic myopathy: biochemical mechanisms. Drug Alcohol Depend. 2201, 63, pp 199–205. https://doi.org/10.1016/s0376-8716(00)00219-2
16. Adachi, J.; Asano, M.; Ueno, Y.; Niemelä, O.; Ohlendieck, K.; Peters, T. J.; Preedy, V. R. Alcoholic muscle disease and biomembrane perturbations (review). J. Nutr. Biochem; 2003, 14 (11), pp 616–625. https://doi.org/10.1016/s0955-2863(03)00114-1
17. Preedy, V. R.; Patel, V. B.; Reilly, M. E.; Richardson, P. J.; Falkous, G.; Mantle, D. Oxidants, antioxidants and alcohol: implications for skeletal and cardiac muscle. Front Biosci; 1999, 4, pp 58–66. https://doi.org/10.2741/a480
18. Vary, T. C.; Nairn, A. C.; Lang, C. H. Restoration of protein synthesis in heart and skeletal muscle after withdrawal of alcohol. Alcohol Clin Exp Res; 2004, 28 (4), pp 517–525. https://doi.org/10.1097/01.alc.0000121653.80502.54
19. Sharma, S. C.; Ray, R. C.; Banerjee, A. K.; Lakshmanan, C. Chronic muscle wasting in alcoholics – a histochemical and biochemical study. Indian J Pathol Microbiol; 1990, 33 (3), pp 244–249.
20. Reilly, M. E.; McKoy, G.; Mantle, D.; Peters, T. J.; Goldspink, G.; Preedy, V. R. Protein and mRNA levels of the myosin heavy chain isoforms I, IIa, IIx and IIb in type I and type II fibrepredominant rat skeletal muscles in response to chronic alcohol feeding. J. Muscle. Res. Cell. Motil; 2000, 21 (8), pp 763–773. https://doi.org/10.1023/a:1010336624154
21. Pipinos, I. I.; Swanson, S. A.; Zhu, Z.; Nella, A. A.; Weiss, D. J.; Gutti, T. L.; McComb, R. D.; Baxter, B. T.; Lynch, T. G.; Casale, G. P. Chronically ischemic mouse skeletal muscle exhibits myopathy in association with mitochondrial dysfunction and oxidative damage. Am J Physiol Regul Integr Comp Physiol; 2008; 295 (1), pp 290–296. https://doi.org/10.1152/ajpregu.90374.2008
22. Slavin, G.; Martin, F.; Ward, P.; Levi, J.; Peters, T. Chronic alcohol excess is associated with selective but reversible injury to type 2B muscle fibers. J Clin Pathol; 1983, 36 (7), pp 772–777. https://doi.org/10.1136/jcp.36.7.772
23. Clary, C. R.; Guidot, D. M.; Bratina, M. A.; Otis, J. S. Chronic alcohol ingestion exacerbates skeletal muscle myopathy in HIV-1 transgenic rats. AIDS Res Ther; 2011, 8 (30), pp 1–9. https://doi.org/10.1186/1742-6405-8-30
24. Barie, P. S.; Mullins, R. J. Experimental methods in the pathogenesis of limb ischemia. J Surg Res; 1988, 44, pp 284–307.
25. Albani, M.; Megalopoulos, A.; Kiskinis, D.; Parashos, S. A.; Grigoriadis, N.; Guiba-Tziampiri, O. Morphological, histochemical, and interstitial pressure changes in the tibialis anterior muscle before and after aortofemoral bypass in patients with peripheral arterial occlusive disease. BMC. Musculoskelet. Disord; 2002, 3 (8), pp 1–7. https://doi.org/10.1186/1471-2474-3-8
26. Rácz, I. B.; Illyés, G.; Sarkadi, L.; Hamar, J. The functional and morphological damage of ischemic reperfused skeletal muscle. Eur Surg Res; 1997, 29 (4), pp 254–263. https://doi.org/10.1159/000129531
27. Ноздренко, Д. М.; Мотузюк, О. П.; Долгополов, О. В.; Заводовський, Д. О. Зміна швидкісно-силових параметрів скорочення скелетних м’язів за умов гострої ішемії. Науковий вісник Волинського національного університету імені Лесі Українки; 2012, 19 (244), c 101–109.
28. Заводовський, Д.; Ноздренко, Д.; Сорока, В.; Хома, О.; Мотузюк, О. Диверсифікація динаміки розвитку втоми ішемізованого м’яза. Науковий вісник Східноєвропейського національного університету імені Лесі Українки. Серія: Біологічні науки; 2014, 13 (290), c 8–17. https://doi.org/10.31558/2617-0248
29. Babinkov, V. І.; Khitrov, N. К.; Cherkashina, Z. A. Effect of early Fascitomy on Intramuscular Pressure and Electrical Excitability of Muscles in Experimental Compartment Syndrome. Bull. Experim. Biol. Med; 2000, 130 (9), pp 857–860.
30. Gordon, A. M.; Homsher, E.; Regnier, M. Regulation of contraction in striated muscle. Physiol Rev; 2000, 80 (2), pp 853–924.https://doi.org/10.1152/physrev.2000.80.2.853
31. Holobar, A.; Farina, D.; Gazzoni, M.; Merletti, R.; Zazula, D. Estimating motor unit discharge patterns from highdensity surface electromyogram. Clin Neurophysiol; 2009, 120 (3), pp 551–562. https://doi.org/10.1016/j.clinph.2008.10.160
32. Vieira, T. M.; Windhorst, U.; Merletti, R. Is the stabilization of quiet upright stance in humans driven by synchronized modulations of the activity of medial and lateral gastrocnemius muscles? J. Appl. Physiol; 2010, 108 (1), pp 85–97.
33. Loerakker, S.; Oomens, C. W.; Manders, E.; Schakel, T.; Bader, D. L.; Baaijens, F. P.; Nicolay, K.; Strijkers, G. J. Ischemia-reperfusion injury in rat skeletal muscle assessed with T2-weighted and dynamic contrast-enhanced MRI. Magn Reson Med; 2011, 66 (2), pp 528–537. https://doi.org/10.1002/mrm.22801
34. Carvalho, A. J.; McKee, N. H.; Green, H. J. Metabolic and contractile responses of fast and slow twitch rat skeletal muscles to ischemia and reperfusion. Plast Reconstr Surg; 1997, 99 (1), pp 163–171. https://doi.org/10.1097/00006534-199701000-00025
35. Vignaud, A.; Hourde, C.; Medja, F.; Agbulut, O.; Butler-Browne, G.; Ferry, A. Impaired skeletal muscle repair after ischemia-reperfusion injury in mice. Journal of Biomedicine and Biotechnology [Online]; 2010, https://dx.doi.org/10.1155% 2F2010%2 F724914 https://doi.org/10.1155/2010/724914
36. Tupling, R.; Green, H.; Senisterra, G.; Lepock, J.; Mckee, N. Effects of ischemia on sarcoplasmic reticulum Са2+ uptake and Са2+ release in rat skeletal muscle. Am J Physiol Endocrinol Metab; 2001, 281, pp 224–232.
37. Tupling, , 2005 A. R. Effects of ischemia and ischemia-reperfusion on sarcoplasmic retilculum. Structure and function in rat sceletal muscle. Thesis for the degree of Doctor of Phisiophy in Kinesiology, University of Waterloo, Canada.
38. Sever, M. S.; Vanholder, R. Crush syndrome: a case report and review of the literature. J Emerg Med; 2015, 48 (6), pp 730–731.
39. Wang, X. T.; Tian, Y.; Xu, W. X.; Cui, L. H.; Xiang, S. Y.; Lü, S. C. Protective effects of modeled superoxide dismutase coordination compound (MSODa) against ischemia/reperfusion injury in rat skeletal muscle. Cell Physiol Biochem; 2015, 37 (2), pp 465–476. https://doi.org/10.1159/000430369
40. Valsoni, B. C. G.; Bonfim, M. R.; Camargo, R. C. T.; Abreu, L. C.; Souza, D. R. S.; Camargo Filho, J. C. S. Effects of Passive Smoking Associated with Physical Exercise in the Skeletal Muscles of Rats During Pregnancy and Lactation. International Journal of Morphology; 2015, 33, pp 497–506. https://doi.org/10.4067/s0717-95022015000200015
41. Brasileiro, J. L.; Fagundes, D. J.; Miiji, L. O. N.; Oshima, C. T. F.; Teruya, R.M.; Guido, I.; Celso M.; Santos, M. A. Isquemia e reperfusão de músculo sóleo de ratos sob ação da pentoxifilina. Jornal Vascular Brasileiro; 2007, 6 (1), pp 50–63. https://dx.doi.org/10.1590/S1677-5449200 7000100008
42. Мальченко, О. А.; Кубышкин, А. В.; Анисимова, Л. В.; Шаланин, В. В.; Мандрик Ю. В. Изменения в мышечной ткани задней конечности крыс в разные сроки формирования синдрома ишемии-реперфузии. Таврический медико-биологический вестник; 2012, 15, (3), ч. 1 (59), с 207–210.
43. Carmo-Araújo, E.M.; Dal-Pai-Silva, M.; Dal-Pai, V.; Cecchini, R.; Anjos Ferreira, A.L. Ischaemia and reperfusion effects on skeletal muscle tissue: morphological and histochemical studies. Int J Exp Pathol; 2007; 88(3), pp 147–154. doi:10.1111/j.1365-2613.2007.00526.x
44. Ohlendieck, K.; Harmon, S.; Koll, M.; Paice, A. G.; Preedy, V. R.; Ca2+-regulatory muscle proteins in the alcohol-fed rat. Metabolism; 2003, 52(9). pp 1102–1112. https://doi.org/10.1016/s0026-0495(03)00063-5
45. Литвина, Н. А. Состояние кровообращения верхних конечностей у больных с диафизарными переломами костей предплечья. Прокопьевск, 1977. 20 с.
46. Chawla, J.; Gruener, G. Management of critical illness polyneuropathy and myopathy. Neurol. Clin; 2010, 28, p 961–977. https://doi.org/10.1016/j.ncl.2010.03.027
47. Gorshkova D. A. Free radical processes state in chronic alcohol intoxication. Actual problems of medicine; 2013, 22, p 191– 193.
48. Hoek, J. B.; Cahill, A.; Pastorino, J. G. Alcohol and Mitochondria: а dysfunction al relationship. Gastroenterol; 2002, 122 (7), p 2049–2063. https://doi.org/10.1053/gast.2002.33613
49. Патченко, Ю. В.; Салютін, Р. В.; Дамбровський, Д. Б. Стан судинного ендотелію та гістологічні зміни м’язової тканини у хворих при хронічній ішемії кінцівок. Клінічна хірургія; 2011, 3, с 41–44.
50. Kauko, A.; Hjelt, L. Morphological changes in striated muscle during ischemia. A clinical and histological study in man. Actaorthop. Scandinav, 1998, 39. p 13–19.
51. Urbano-Marquez, A.; Fernandez-Sola, J. The effects of alcohol on skeletal and cardiac muscle. Muscle Nerve; 2004, 30. p 689–707. https://doi.org/10.1002/mus.20168



How to Cite

Motuziuk, O., & Pykaliuk, V. (2019). Histologic Features of Rat Gastrocnemius Muscle Under Conditions of Experimental Ischemia of Different Duration Complicated by Chronic Alcoholization. Notes in Current Biology, (3(387), 137–147. https://doi.org/10.29038/2617-4723-2019-387-137-147