The Effect of Resistance Training with Boldenone Consumption, Ziziphus jujuba Extract and Gallic Acid on Annexin-5 and Caspase-3 Genes Expression in Cardiac Tissue of Male Wistar Rats

Document Type : Original Article


Department of Exercise Physiology, Ayatollah Amoli Branch, Islamic Azad University, Amol, Iran


Introduction: Herbal medicines have gained growing popularity and have been used worldwide as alternative and complementary medicine and food supplements. The aim of this study was to investigate the effect of resistance training with boldenone consumption, ziziphus jujuba extract and Gallic acid on annexin-5 and caspase-3 genes expression in cardiac tissue of male Wistar rats. Methods: In this experimental study, 42 male Wistar rats aged 8-12 weeks with mean weight of 195±7.94g were randomly divided into seven groups: control, boldenone, jujube extract + boldenone, Gallic acid + boldenone, resistance training + boldenone, resistance training + jujube extract + boldenone, resistance training + Gallic acid + boldenone. Resistance training program included six weeks of climbing a ladder for 5 sessions, 3 sets, 5 repetitions and one-minute resting between sets with 50% 1RM in the first week and 100% 1RM in the final week. The drug was injected in the hamstring in depth once a week on an appointed day. After anesthesia, the autopsy was performed and cardiac tissue was dissected. Data were analyzed using paired sample t-test, One-way ANOVA and Scheffe post hoc at the P


1. Cabral AM, Vasquez EC, Moysés MR, Antonio A. Sex hormone modulation of ventricular hypertrophy in sinoaortic denervated rats. Hypertension. 1988; 11 (22): I93.
2. Malhotra A, Buttrick P, Scheuer J. Effects of sex hormones on development of physiological and pathological cardiac hypertrophy in male and female rats. Am J Physiol. 1990; 259 (3): 866- 871.
3. Payne J, Kotwinski P, Montgomery H. Cardiac effects of anabolic steroids. Heart. 2004; 90 (5): 473- 475.
4. Hassan AF, Kamal MM. Effect of exercise training and anabolic androgenic steroids on hemodynamics, glycogen content, angiogenesis and apoptosis of cardiac muscle in adult male rats. International journal of health sciences. Int J Health Sci. 2013; 7 (1): 47- 60.
5. Fineschi V, Riezzo I, Centini F, Silingardi E, Licata M, Beduschi G, et al. Sudden cardiac death during anabolic steroid abuse: morphologic and toxicologic findings in two fatal cases of bodybuilders. Int J Legal Med. 2007; 121 (1): 48- 53.
6. Sgoifo Rossi C, Arioli F, Bassini A, Chiesa L, Dell'Orto V, Montana M, et al. Evidence for false-positive results for boldenone testing of veal urine due to faecal cross-contamination during sampling. Food Addit Contam. 2004; 21 (8): 756- 762.
7. Pinho CA, Tromm CB, Tavares AM, Silva LA, Silveira PCL, Souza CT, et al. Effects of different physical training protocols on ventricular oxidative stress parameters in infarction-induced rats. Life Sci. 2012; 90 (13): 553- 559.
8. Leosco D, Rengo G, Iaccarino G, Golino L, Marchese M, Fortunato F, et al. Exercise promotes angiogenesis and improves β-adrenergic receptor signalling in the post-ischaemic failing rat heart. Cardiovasc Res. 2008; 78 (2): 385- 394.
9. Beavers KM, Brinkley TE, Nicklas BJ. Effect of exercise training on chronic inflammation. Clin Chim Acta. 2010; 411 (11): 785- 793.
10. Huang CY, Yang AL, Lin YM, Wu FN, Lin JA, Chan YS, et al. Anti-apoptotic and pro-survival effects of exercise training on hypertensive hearts. J Appl Physiol. 2012; 112 (5): 883- 891.
11. Santana ET, Serra AJ, Silva Junior JA, Bocalini DS, Barauna VG, Krieger JE, et al. Aerobic exercise training induces an anti-apoptotic milieu in myocardial tissue. Motriz Rev Educ Fis. 2014; 20 (2): 233- 238.
12. Siu PM, Bryner RW, Martyn JK, Alway SE. Apoptotic adaptations from exercise training in skeletal and cardiac muscles. FASEB J. 2004; 18 (10): 1150- 1152.
13. Thompson CB. Apoptosis in the pathogenesis and treatment of disease. Science. 1995; 267 (5203): 1456.
14. Shokri S, Aitken RJ, Abdolvahhabi M, Abolhasani F, Ghasemi FM, Kashani I, et al. Exercise and supraphysiological dose of nandrolone deconoate increase apoptosis in spermatogenic cells. Basic Clin Pharmacol Toxicol. 2010; 106 (4): 324- 330.
15. Condorelli G, Roncarati R, Ross J, Pisani A, Stassi G, Todaro M, et al. Heart-targeted overexpression of caspase3 in mice increases infarct size and depresses cardiac function. Proc Natl Acad Sci. 2001; 98 (17): 9977- 9982.
16. Narula J, Haider N, Arbustini E, Chandrashekhar Y. Mechanisms of disease: apoptosis in heart failure—seeing hope in death. Nat Clin Pract Cardiovasc Med. 2006; 3 (12): 681- 688.
17. Laohavisit A, Davies JM. Multifunctional annexins. Plant Sci. 2009; 177 (6): 532- 539.
18. Van Genderen H, Kenis H, Lux P, Ungeth L, Maassen C, Deckers N, et al. In vitro measurement of cell death with the annexin A5 affinity assay. Nat Protoc. 2006; 1 (1): 363.
19. Hofstra L, Liem H, Dumont EA, Boersma HH, van Heerde WL, Doevendans PA, et al. Visualisation of cell death in vivo in patients with acute myocardial infarction. Lancet. 2000; 356 (9225): 209- 212.
20. Haykowsky MJ. Left ventricular remodelling and the athlete's heart: time to revisit the Morganroth hypothesis. J Physiol. 2011; 589 (24): 5915- 5919.
21. Barauna VG, Rosa KT, Irigoyen MC, De Oliveira EM. Effects of resistance training on ventricular function and hypertrophy in a rat model. Clin Med Res. 2007; 5 (2): 114- 120.
22. Doustar Y, Soufi FG, Saber MM, Ghiassie R, Jafary A. Role of four-week resistance exercise in preserving the heart against ischaemia-reperfusion-induced injury: cardiovascular topic. Cardiovasc J Afr. 2012; 23 (8): 451- 455.
23. Kwak HB, Song W, Lawler JM. Exercise training attenuates age-induced elevation in Bax/Bcl-2 ratio, apoptosis, and remodeling in the rat heart. FASEB J. 2006; 20 (6): 791- 793.
24. Riezzo I, Di Paolo M, Neri M, Bello S, Cantatore S, D'Errico S, et al. Anabolic steroid-and exercise-induced cardio-depressant cytokines and myocardial β1 receptor expression in CD1 mice. Curr Pharm Biotechnol. 2011; 12 (2): 275- 284.
25. Ebrahimi S, Sadeghi H, Pourmahmoudi A, Askariyan S, Askari S. Protective effect of zizphus vulgaris extract, on liver toxicity in laboratory rats. Armaghane Danesh J. 2011; 16 (2): 172- 180.
26. Al-Reza SM, Bajpai VK, Kang SC. Antioxidant and antilisterial effect of seed essential oil and organic extracts from Zizyphus jujuba. Food Chem Toxicol. 2009; 47 (9): 2374- 2380.
27. Li JW, Fan LP, Ding SD, Ding XL. Nutritional composition of five cultivars of Chinese jujube. Food Chem. 2007; 103 (2): 454- 460.
28. Benammar C, Hichami A, Yessoufou A, Simonin A-M, Belarbi M, Allali H, et al.
Zizyphus lotus L.(Desf.) modulates antioxidant activity and human T-cell proliferation. ISCMR. 2010; 10 (1): 54- 63.
29. Mesulam MM, Guillozet A, Shaw P, Levey A, Duysen E, Lockridge O. Acetylcholinesterase knockouts establish central cholinergic pathways and can use butyrylcholinesterase to hydrolyze acetylcholine. Neuroscience. 2002; 110 (4): 627- 639.
30. Sun YF, Song CK, Viernstein H, Unger F, Liang ZS. Apoptosis of human breast cancer cells induced by microencapsulated betulinic acid from sour jujube fruits through the mitochondria transduction pathway. Food Chem. 2013; 138 (2): 1998- 2007.
31. Plastina P, Bonofiglio D, Vizza D, Fazio A, Rovito D, Giordano C, et al. Identification of bioactive constituents of Ziziphus jujube fruit extracts exerting antiproliferative and apoptotic effects in human breast cancer cells. J Ethnopharmacol. 2012; 140 (2): 325- 332.
32. Abedini MR, Erfanian N, Nazem H, Jamali S, Hoshyar R. Anti-proliferative and apoptotic effects of Ziziphus Jujube on cervical and breast cancer cells. Avicenna J Phytomed. 2016; 6 (2): 142.
33. Niaki AG, Hosseini F, Rooadbari F, Ahmadabad SR, Rooadbari M. Effects of aerobic training, with or without Zizyphus jujuba water extraction, on fundus nesfatin-1, ATP, HDL-C, and LDL-C concentrations in female rats. J Phys Activ Health. 2013; 4 (1): 9- 16.
34. Kaur M, Velmurugan B, Rajamanickam S, Agarwal R, Agarwal C. Gallic acid, an active constituent of grape seed extract, exhibits anti-proliferative, pro-apoptotic and anti-tumorigenic effects against prostate carcinoma xenograft growth in nude mice. Pharm Res. 2009; 26 (9): 2133- 2140.
35. Liu Z, Li D, Yu L, Niu F. Gallic acid as a cancer-selective agent induces apoptosis in pancreatic cancer cells. Chemotherapy. 2012; 58 (3): 185- 194.
36. Lee S, Farrar RP. Resistance training induces muscle-specific changes in muscle mass and function in rat. J Exerc Physiol Online. 2003; 6 (2): 80- 87.
37. Rosano G, Cornoldi A, Fini M. Effects of androgens on the cardiovascular system. J Endocrinol Invest. 2004; 28 (3): 32- 38.
38. Zaugg M, Jamali NZ, Lucchinetti E, Xu W, Alam M, Shafiq SA, et al. Anabolic‐androgenic steroids induce apoptotic cell death in adult rat ventricular myocytes. J Cell Physiol. 2001; 187 (1): 90- 95.
39. Lieberherr M, Grosse B. Androgens increase intracellular calcium concentration and inositol 1, 4, 5-trisphosphate and diacylglycerol formation via a pertussis toxin-sensitive G-protein. J Biol Chem. 1994; 269 (10): 7217- 7223.
40. Kroemer G, Dallaporta B, Resche-Rigon M. The mitochondrial death/life regulator in apoptosis and necrosis. Annu Rev Physiol. 1998; 60 (1): 619- 642.
41. Ellison GM, Waring CD, Vicinanza C, Torella D. Physiological cardiac remodelling in response to endurance exercise training: cellular and molecular mechanisms. Heart. 2011; 30063- 30069.
42. Riezzo I, De Carlo D, Neri M, Nieddu A, Turillazzi E, Fineschi V. Heart disease induced by AAS abuse, using experimental mice/rats models and the role of exercise-induced cardiotoxicity. Mini Rev Med Chem. 2011; 11 (5): 409- 424.
43. Fanton L, Belhani D, Vaillant F, Tabib A, Gomez L, Descotes J, et al. Heart lesions associated with anabolic steroid abuse: comparison of post-mortem findings in athletes and norethandrolone-induced lesions in rabbits. Exp Toxicol Pathol. 2009; 61 (4): 317- 323.
44. Belhani D, Fanton L, Vaillant F, Descotes J, Manati W, Tabib A, et al. Cardiac lesions induced by testosterone: protective effects of dexrazoxane and trimetazidine. Cardiovasc Toxicol. 2009; 9 (2): 64- 69.