تأثیر تعاملی تمرین هوازی در آب و مکمل کروسین بر بیان ژن‌های BAX و BCL-2 کاردیومایوسیتی در رت‌های نر تیمار شده با پراکسید هیدروژن

نوع مقاله : مقاله های پژوهشی

نویسندگان

1 دانشجوی دکتری تخصصی، گروه فیزیولوژی ورزشی، دانشکده‌ی علوم ورزشی، دانشگاه تربیت دبیر شهید رجایی، تهران، ایران

2 استادیار، گروه فیزیولوژی ورزشی، دانشکده‌ی علوم ورزشی، دانشگاه تربیت دبیر شهید رجایی، تهران، ایران

3 دانشیار، گروه فیزیولوژی ورزشی، دانشکده تربیت بدنی و علوم ورزشی، دانشگاه خوارزمی، تهران، ایران

4 استاد، گروه فیزیولوژی ورزشی، دانشکده‌ی علوم ورزشی، دانشگاه تربیت دبیر شهید رجایی، تهران، ایران

5 دکتری تخصصی، مرکز تحقیقات علوم پایه، شرکت هیستوژنوتک، تهران، ایران

چکیده

مقدمه: هدف از انجام پژوهش حاضر، تعیین اثر تعامل تمرین هوازی در آب و مکمل کروسین بر بیان ژن‌های BAX و BCL-2 کاردیومایوسیتی در رت‌های نر مسموم شده با 2O2H بود.روش‌ها: 36 سر رت‌ جوان نژاد ویستار با میانگین وزنی 180 تا 200 گرم به صورت تصادفی به شش گروه «شم، 2O2H، 2O2H + کروسین، 2O2H + فعالیت در آب، 2O2H + فعالیت در آب + کروسین و گروه شاهد» تقسیم شدند. فعالیت در آب 5 روز در هفته دقیقه، هر بار 60 دقیقه و به مدت شش هفته صورت گرفت. 1 میلی‌مول بر کیلوگرم 2O2H و 5/12 میلی‌مول بر کیلوگرم کروسین نیز به صورت داخل صفاقی تزریق گردید. به منظور بیان ژن‌ها از دستگاه Real time polymerase chain reaction (Real Time PCR) و برای تأیید بروز آپوپتوز از روش Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) استفاده شد. جهت تجزیه و تحلیل داده‌ها آزمون‌های Kolmogorov-Smirnov، Levene، One-Way ANOVA و Scheffe مورد استفاده قرار گرفت.یافته‌ها: القای 2O2H با افزایش معنی‌دار بیان BAX همراه بود (001/0 = P). شش هفته فعالیت در آب منجر به کاهش بیان BCL-2 (023/0 = P) و القای کروسین نیز باعث کاهش معنی‌دار بیان BAX شد. تعامل بین فعالیت‌های القای کروسین و القای 2O2H اثر کاهنده‌ای بر بیان BAX و اثر افزاینده‌ای بر بیان BCL-2 داشت که نشان دهنده‌ی تأثیر حفاظتی فعالیت منظم در کاهش آپوپتوز کاردیومایوسیتی می‌باشد.نتیجه‌گیری: فعالیت منظم در آب و همچنین، مصرف کروسین به صورت جداگانه، منجر به کاهش معنی‌داری در بیان BAX و افزایش معنی‌داری در بیان BCL-2 می‌شود. از سوی دیگر، تلفیق فعالیت منظم و کروسین تأثیر هم‌افزایی و سینرژیک در مهار بیان BAX و افزایش بیان BCL-2 در رت‌های مسموم شده با 2O2H دارد

کلیدواژه‌ها


عنوان مقاله [English]

The Interactive Effect of Forced Swimming and Crocin Supplementation on the Expression of BAX and BCL-2 Cardiomyocyte Genes in Male Rats Infected with Hydrogen Peroxide

نویسندگان [English]

  • Mina Akbari 1
  • Fereshteh Shahidi 2
  • Hamid Rajabi 3
  • Majid Kashef 4
  • Zohreh Mazaheri 5
1 PhD Student, Department of Exercise Physiology, School of Sport Sciences, Shahid Rajaee Teacher Training University, Tehran, Iran
2 Assistant Professor, Department of Exercise Physiology, School of Sport Sciences, Shahid Rajaee Teacher Training University, Tehran, Iran
3 Associate Professor, Department of Exercise Physiology, School of Physical Education and Sport Sciences, Kharazmi University, Tehran, Iran
4 Professor, Department of Exercise Physiology, School of Sport Sciences, Shahid Rajaee Teacher Training University, Tehran, Iran
5 PhD, Basic Medical Science Research Center, Histogenotech Companey, Tehran, Iran
چکیده [English]

Background: The aim of this study was to determine the interaction effect of forced swimming and crocin supplementation on the expression of BAX and BCL-2 cardiomyocyte genes in male rats poisoned by hydrogen peroxide (H2O2).Methods: 36 male Wistar rats were randomly divided into six groups of sham, H2O2, H2O2 and crocin, H2O2 and forced swimming, H2O2 and crocin and forced swimming, and control. The forced swimming protocol was 60 minutes, 5 days a week, for 6 weeks. H2O2 (1 mg/kg for 3 weeks) and crocin (12.5 Mm/kg daily with 5 cc of distilled water) were injected intraperitoneally. Real time polymerase chain reaction (PCR) device was used to assess gene expression, and apoptosis was assessed using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method. Statistical analysis was done using Kolmogorov-Smirnov, Levene, one-way ANOVA and, the Scheffe post hoc tests.Findings: H2O2 induction was associated with a significant increase in the apoptosis of BAX (P < 0.001). Six weeks of exercise in water tended to reduction in the BCL-2 expression (P = 0.023), and induction of crocin resulted in significant decrease in BAX expression. The interaction between swimming, crocin and H2O2 decreased BAX and increased BCL-2 expression, which indicated the protective effect of regular activity in reducing cardiomyocyte apoptosis.Conclusion: Forced swimming as well as the use of crocin separately led to a significant reduction in BAX gene expression and a significant increase in BCL-2 gene expression. On the other hand, the combination of exercise and crocin had synergistic and synergistic effects in inhibiting the apoptotic expression of BAX and also enhancing the expression of the anti-apoptotic gene of BCL-2 in rats poisoned by H2O2.

کلیدواژه‌ها [English]

  • Forced swimming
  • Crocin
  • BAX protein
  • BCL2 proteins
  • Hydrogen peroxide
  • Cardiomyocyte
  1. Halliwell B, Gutteridge J. Free Radicals in Biology and Medicine. New York, NY: Oxford University Press; 2015.
  2. Persson T, Popescu BO, Cedazo-Minguez A. Oxidative stress in Alzheimer's disease: Why did antioxidant therapy fail? Oxid Med Cell Longev 2014; 2014: 427318.
  3. Valko M, Jomova K, Rhodes CJ, Kuca K, Musilek K. Redox- and non-redox-metal-induced formation of free radicals and their role in human disease. Arch Toxicol 2016; 90(1): 1-37.
  4. Kelly FJ. Oxidative stress: its role in air pollution and adverse health effects. Occup Environ Med 2003; 60(8): 612-6.
  5. Faria A, Persaud SJ. Cardiac oxidative stress in diabetes: Mechanisms and therapeutic potential. Pharmacol Ther 2017; 172: 50-62.
  6. Pisoschi AM, Pop A. The role of antioxidants in the chemistry of oxidative stress: A review. Eur J Med Chem 2015; 97: 55-74.
  7. Lajoie C, Calderone A, Beliveau L. Exercise training enhanced the expression of myocardial proteins related to cell protection in spontaneously hypertensive rats. Pflugers Arch 2004; 449(1): 26-32.
  8. Oruc S, Gonul Y, Tunay K, Oruc OA, Bozkurt MF, Karavelioglu E, et al. The antioxidant and antiapoptotic effects of crocin pretreatment on global cerebral ischemia reperfusion injury induced by four vessels occlusion in rats. Life Sci 2016; 154: 79-86.
  9. Ascensao A, Ferreira R, Magalhaes J. Exercise-induced cardioprotection--biochemical, morphological and functional evidence in whole tissue and isolated mitochondria. Int J Cardiol 2007; 117(1): 16-30.
  10. Mohsenizadeh N, Azarbayjani MA, Najafipour H, Matin Homaee H, Keshtkar A. The simultaneous effect of regular exercise and vitamin d on NF-kBp65 levels in male rats exposed to hydrogen peroxide. Knowledge Health 2017; 12(3): 55-62. [In Persian].
  11. Granger DN, Kvietys PR. Reperfusion injury and reactive oxygen species: The evolution of a concept. Redox Biol 2015; 6: 524-51.
  12. Roy S, Nicholson DW. Cross-talk in cell death signaling. J Exp Med 2000; 192(8): F21-F25.
  13. Ghavami S, Hashemi M, Kadkhoda K, Alavian SM, Bay GH, Los M. Apoptosis in liver diseases--detection and therapeutic applications. Med Sci Monit 2005; 11(11): RA337-RA345.
  14. Li PF, Dietz R, von Harsdorf R. p53 regulates mitochondrial membrane potential through reactive oxygen species and induces cytochrome c-independent apoptosis blocked by Bcl-2. EMBO J 1999; 18(21): 6027-36.
  15. Janero DR, Hreniuk D, Sharif HM. Hydrogen peroxide-induced oxidative stress to the mammalian heart-muscle cell (cardiomyocyte): Lethal peroxidative membrane injury. J Cell Physiol 1991; 149(3): 347-64.
  16. Radak Z, Sasvari M, Nyakas C, Pucsok J, Nakamoto H, Goto S. Exercise preconditioning against hydrogen peroxide-induced oxidative damage in proteins of rat myocardium. Arch Biochem Biophys 2000; 376(2): 248-51.
  17. Sallam N, Laher I. Exercise modulates oxidative stress and inflammation in aging and cardiovascular diseases. Oxid Med Cell Longev 2016; 2016: 7239639.
  18. 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): 791-3.
  19. Melo SF, Fernandes T, Barauna VG, Matos KC, Santos AA, Tucci PJ, et al. Expression of microRNA-29 and Collagen in cardiac muscle after swimming training in myocardial-infarcted rats. Cell Physiol Biochem 2014; 33(3): 657-69.
  20. Aruoma OI. Free radicals, oxidative stress, and antioxidants in human health and disease. J Amer Oil Chem Soc 1998; 75(2): 199-212.
  21. Verma SK, Bordia A. Antioxidant property of Saffron in man. Indian J Med Sci 1998; 52(5): 205-7.
  22. Sachdeva J, Tanwar V, Golechha M, Siddiqui KM, Nag TC, Ray R, et al. Crocus sativus L. (saffron) attenuates isoproterenol-induced myocardial injury via preserving cardiac functions and strengthening antioxidant defense system. Exp Toxicol Pathol 2012; 64(6): 557-64.
  23. Mehdizadeh R, Parizadeh MR, Khooei AR, Mehri S, Hosseinzadeh H. Cardioprotective effect of saffron extract and safranal in isoproterenol-induced myocardial infarction in wistar rats. Iran J Basic Med Sci 2013; 16(1): 56-63.
  24. Razavi BM, Imenshahidi M, Abnous K, Hosseinzadeh H. Cardiovascular effects of saffron and its active constituents: A review article. Saffron agronomy and technology 2014; 1(2): 3-13. [In Persian].
  25. Arshadi S, Azarbayjani MA, Hajaghaalipor F, Yusof A, Peeri M, Bakhtiyari S, et al. Evaluation of Trigonella foenum-graecum extract in combination with swimming exercise compared to glibenclamide consumption on type 2 Diabetic rodents. Food Nutr Res 2015; 59: 29717.
  26. Radak Z, Sasvari M, Nyakas C, Pucsok J, Nakamoto H, Goto S. Exercise preconditioning against hydrogen peroxide-induced oxidative damage in proteins of rat myocardium. Arch Biochem Biophys 2000; 376(2): 248-51.
  27. Lari P, Rashedinia M, Abnous K, Hosseinzadeh H. Crocin improves lipid dysregulation in subacute diazinon exposure through ERK1/2 pathway in rat liver. Drug Res (Stuttg) 2014; 64(6): 301-5.
  28. Maletic SD, Dragicevic-Djokovic LM, Ognjanovic BI, Zikic RV. Alterations of rat reticulocyte (anti)oxidant status and energy metabolism influenced by hydrogen-peroxide [1999]. Acta Biologica Iugoslavica 2004; 35(3): 129-38.
  29. Cheng M, Chen Y, Xiao W, Sun R, Tian Z. NK cell-based immunotherapy for malignant diseases. Cell Mol Immunol 2013; 10(3): 230-52.
  30. Siu PM, Bryner RW, Martyn JK, Alway SE. Apoptotic adaptations from exercise training in skeletal and cardiac muscles. FASEB J 2004; 18(10): 1150-2.
  31. Xiang M, Qian ZY, Zhou CH, Liu J, Li WN. Crocetin inhibits leukocyte adherence to vascular endothelial cells induced by AGEs. J Ethnopharmacol 2006; 107(1): 25-31.
  32. Xu G, Gong Z, Yu W, Gao L, He S, Qian Z. Increased expression ratio of Bcl-2/Bax is associated with crocin-mediated apoptosis in bovine aortic endothelial cells. Basic Clin Pharmacol Toxicol 2007; 100(1): 31-5.
  33. Xu G, Gong Z, Yu W, Gao L, He S, Qian Z. Increased expression ratio of Bcl-2/Bax is associated with crocin-mediated apoptosis in bovine aortic endothelial cells. Basic Clin Pharmacol Toxicol 2007; 100(1): 31-5.
  34. Xiang M, Yang M, Zhou C, Liu J, Li W, Qian Z. Crocetin prevents AGEs-induced vascular endothelial cell apoptosis. Pharmacol Res 2006; 54(4): 268-74.
  35. Varmazyar M, Azarbayjani M. The effect of saffron supplementation of antioxidant enzymes activities during a session eccentric exercise in active Males. J Med Plants 2014; 2(50): 54-63. [In Persian].