Effect of Aerobic Training and Eugenol on the Expression of SERCA2a and NKX2-5 Genes in Heart Tissue of Rats Poisoned with Chlorpyrifos

Document Type : Research Paper

Authors

1 Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University, Tehran, Iran

2 Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University, Tehran, Iran.

3 Department of Physical Education, Pardis Branch, Islamic Azad University, Pardis, Iran.

Abstract
Chlorpyrifos is an organophosphate toxin used to control agricultural pests. These toxins cause oxidative stress and apoptosis in various tissues of the body, including the heart. SERCA2a and NKX2-5 are genes that regulate contractile function and heart growth. Previous studies have shown a positive effect of physical activity on the expression of these genes in heart failure, but so far, changes in these genes following the use of physical activity in chlorpyrifos poisoning have not been studied. The aim of the present study was to investigate the independent and interactive effect of aerobic exercise and eugenol supplementation on the expression of SERCA2a and NKX2-5 genes in the heart tissue of rats poisoned with chlorpyrifos. Thirty-six Wistar male rats were divided into six groups (six in each group) with the names healthy control, sham, poison control, aerobic exercise poison, eugenol poison, and toxic + aerobic exercise + eugenol. Aerobic training was performed on a rodent treadmill (five sessions per week, each session lasting 20 minutes at a speed of 26 meters per minute) for four weeks. Eugenol at a dose of 250 mg/kg body weight was fed to the rats of the supplement groups (five days a week for four weeks using gavage). Data were analyzed using one-way ANOVA with Tukey post-hoc test and P <0.05 was considered as a significant change. The results of the current study showed that poisoning significantly reduced the expression of SERCA2a and NKX2-5 genes in cardiac tissue. Exercise (no supplementation) increased the SERCA2a and NKX2-5 genes in the hearts of poisoned rats. Exercise-eugenol interaction did not cause a synergistic effect. Overall, in the present study, exercise seems to be a more important factor than eugenol in reducing the effect of chronic intoxication on the heart. Further studies with higher doses of eugenol supplementation are needed.

Keywords

  1. Joko T, Dewanti NAY, Dangiran HL. Pesticide poisoning and the use of personal protective equipment (PPE) in Indonesian farmers. Journal of Environmental and Public Health. 2020;2020:5379619.
  2. Sidhu GK, Singh S, Kumar V, Dhanjal DS, Datta S, Singh J. Toxicity, monitoring and biodegradation of organophosphate pesticides: a review. Critical Reviews in Environmental Science and Technology. 2019;49(13):1135-87.
  3. González-Alzaga B, Romero-Molina D, López-Flores I, Giménez-Asensio MJ, Hernández AF, Lacasaña M. Urinary levels of organophosphate pesticides and predictors of exposure in pre-school and school children living in agricultural and urban communities from south Spain. Environmental Research. 2020;186:109459.
  4. Reddy BS, Skaria TG, Polepalli S, Vidyasagar S, Rao M, Kunhikatta V, Nair S, Thunga G. Factors associated with outcomes in organophosphate and carbamate poisoning: a retrospective study. Toxicol Res. 2020; 36(3):257-66.
  5. Roth A, Zellinger I, Arad M, Atsmon J. Organophosphates and the Heart. CHEST. 1993;103(2):576-82.
  6. Zhihao L, Jingyu N, Lan L, Michael S, Rui G, Xiyun B, et al. SERCA2a: a key protein in the Ca(2+) cycle of the heart failure. Heart Fail Rev. 2020;25(3):523-35.
  7. Lipskaia L, Chemaly ER, Hadri L, Lompre A-M, Hajjar RJ. Sarcoplasmic reticulum Ca(2+) ATPase as a therapeutic target for heart failure. Expert Opinion on Biological Therapy. 2010;10(1):29-41.
  8. George V, Colombo S, Targoff KL. An early requirement for nkx2.5 ensures the first and second heart field ventricular identity and cardiac function into adulthood. Dev Biol. 2015;400(1):10-22.
  9. Ueyama T, Kasahara H, Ishiwata T, Nie Q, Izumo S. Myocardin expression is regulated by Nkx2.5, and its function is required for cardiomyogenesis. Mol Cell Biol. 2003;23(24):9222-32.
  10. Anderson DJ, Kaplan DI, Bell KM, Koutsis K, Haynes JM, Mills RJ, et al. NKX2-5 regulates human cardiomyogenesis via a HEY2 dependent transcriptional network. Nature Communications. 2018; 9(1):1373.
  11. Akazawa H, Komuro I. Cardiac transcription factor Csx/Nkx2-5: its role in cardiac development and diseases. Pharmacol Ther. 2005;107(2):252-68.
  12. Ojha A, Yaduvanshi SK, Srivastava N. Effect of combined exposure of commonly used organophosphate pesticides on lipid peroxidation and antioxidant enzymes in rat tissues. Pesticide Biochemistry and Physiology. 2011;99(2):148-56.
  13. Smith EG, Gordon CJ. The effects of chlorpyrifos on blood pressure and temperature regulation in spontaneously hypertensive rats. Basic Clin Pharmacol Toxicol. 2005;96(6):503-11.
  14. Azza ME, El-Sayed ME, Nashwa MB, Afaf MA, Abdelaziz MH, Intisar IA. Cardiovascular toxic effects of chlorpyrifos: a possible protective role for pomegranate extracts. J Clin Toxicol. 2018;8(1):1000374.
  15. Cetin N, Cetin E, Eraslan G, and Bilgili A. Chlorpyrifos induces cardiac dysfunction in rabbits. Res Vet Sci. 2007;82(3):405-8.
  16. Tuzmen N, Candan N, Kaya E, Demiryas N. Biochemical effects of chlorpyrifos and deltamethrin on altered antioxidative defense mechanisms and lipid peroxidation in rat liver. Cell Biochem Funct. 2008;26(1):119-24.
  17. Shirvani H, Ghanbari-Niaki A, Rahmati-Ahmadabad S, Sobhani VJIJoAEP. Effects of endurance training and herb supplementation on tissue nesfatin-1/nucleobindin-2 and ghrelin mRNA expression. Iinternational Journal of Applied Exercise Phusilogy. 2017;6(1):72-85.
  18. Rahmati-Ahmadabad S, Azarbayjani M-A, Broom DR, and Nasehi M. Effects of high-intensity interval training and flaxseed oil supplement on learning, memory and immobility: relationship with BDNF and TrkB genes. Comparative Exercise Physiology. 2021;17(3):273-83.
  19. Zarezadehmehrizi A, Hong J, Lee J, Rajabi H, Gharakhanlu R, Naghdi N, et al. Exercise training ameliorates cognitive dysfunction in amyloid beta-injected rat model: possible mechanisms of Angiostatin/VEGF signaling. Metab Brain Dis. 2021.
  20. Nikbin S, Derakhshideh A, Hozouri Tarighe M, Khojasteh Z, Kanozi F, Mousavi N, et al. Synergic effects of aerobic exercise and eugenol supplement on germ cell development and testicular tissue structure in chlorpyrifos-treated animal model. Environ Sci Pollut Res Int. 2020;27(14):17229-42.
  21. Nikbin S, Tajik A, Allahyari P, Matin G, Hoseini Roote SS, Barati E, et al. Aerobic exercise and eugenol supplementation ameliorated liver injury induced by chlorpyrifos via modulation acetylcholinesterase activation and antioxidant defense. Environmental Toxicology. 2020;35(7):783-93.
  22. Jaganathan SK, Mazumdar A, Mondhe D, Mandal M. Apoptotic effect of eugenol in human colon cancer cell lines. Cell Biol Int. 2011;35(6):607-15.
  23. Júnior PL, Câmara DA, Costa AS, Ruiz JL, Levy D, Azevedo RA, et al. Apoptotic effect of eugenol envolves G2/M phase abrogation accompanied by mitochondrial damage and clastogenic effect on cancer cell in vitro. Phytomedicine. 2016;23(7):725-35.
  24. Al-Sharif I, Remmal A, Aboussekhra A. Eugenol triggers apoptosis in breast cancer cells through E2F1/survivin down-regulation. BMC Cancer. 2013;13:600.
  25. Wisløff U, Loennechen JP, Currie S, Smith GL, Ellingsen Ø. Aerobic exercise reduces cardiomyocyte hypertrophy and increases contractility, Ca2+ sensitivity and SERCA2 in rat after myocardial infarction. Cardiovasc Res. 2002;54(1):162-74.
  26. Morissette MP, Susser SE, Stammers AN, Moffatt TL, Wigle JT, Wigle TJ, et al. Exercise-induced increases in the expression and activity of cardiac sarcoplasmic reticulum calcium ATPase 2 is attenuated in AMPKα(2) kinase-dead mice. Can J Physiol Pharmacol. 2019;97(8):786-95.
  27. Melo SFS, Barauna VG, Neves VJ, Fernandes T, Lara LdS, Mazzotti DR, et al. Exercise training restores the cardiac microRNA-1 and -214 levels regulating Ca2+ handling after myocardial infarction. BMC Cardiovascular Disorders. 2015;15:166-6.
  28. Naderi N, Hemmatinafar M, Gaeini AA, Bahramian A, Ghardashi-Afousi A, Kordi MR, et al. High-intensity interval training increase GATA4, CITED4 and c-Kit and decreases C/EBPβ in rats after myocardial infarction. Life Sciences. 2019;221:319-26.
  29. Zhou X, Xu M, Bryant JL, Ma J, Xu X. Exercise-induced myokine FNDC5/irisin functions in cardiovascular protection and intracerebral retrieval of synaptic plasticity. Cell & Bioscience. 2019;9(1):32.
  30. Seo H, Li HY, Perez-Reyes E, Lee JH. Effects of eugenol on T-type Ca2+ channel isoforms. J Pharmacol Exp Ther. 2013;347(2):310-7.
  31. Nikbin S, Derakhshideh A, Karimi Jafari S, Mirzahamedani A, Moslehi A, Ourzamani S, et al. Investigating the protective effect of aerobic exercise on oxidative stress and histological damages of testicular tissue associated with chlorpyrifos in male rats. Andrologia. 2020;52(2):e13468.
  32. Singh P, Jayaramaiah RH, Agawane SB, Vannuruswamy G, Korwar AM, Anand A, et al. Potential dual role of eugenol in inhibiting advanced glycation end products in diabetes: proteomic and mechanistic insights. Scientific Reports. 2016; 6(1):18798.
  33. Rahmati-Ahmadabad S, Azarbayjani MA, Farzanegi P, Moradi L. High-intensity interval training has a greater effect on reverse cholesterol transport elements compared with moderate-intensity continuous training in obese male rats. Eur J Prev Cardiol. 2019:2047487319887828.
  34. Livak KJ Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25(4):402-8.
  35. Fang B, Li JW, Zhang M, Ren FZ, Pang GF. Chronic chlorpyrifos exposure elicits diet-specific effects on metabolism and the gut microbiome in rats. Food Chem Toxicol. 2018;111:144-52.
  36. Zhang Y, Chang Y, Cao H, Xu W, Li Z, Tao L. Potential threat of Chlorpyrifos to human liver cells via the caspase-dependent mitochondrial pathways. Food and Agricultural Immunology. 2018;29(1):294-305.
  37. Bersaoui M, Baldew SM, Cornelis N, Toelsie J, Cornelissen VA. The effect of exercise training on blood pressure in African and Asian populations: a systematic review and meta-analysis of randomized controlled trials. Eur J Prev Cardiol. 2020; 27(5):457-72.
  38. Herring MP, Puetz TW, O'Connor PJ, Dishman RK. Effect of exercise training on depressive symptoms among patients with a chronic illness: a systematic review and meta-analysis of randomized controlled trials. Arch Intern Med. 2012;172(2):101-11.
  39. Frajtabar Z, Fathi R, Nasiri K, Ahmadi F. The effect of aerobic exercise and ethanol consumption on nrf2 gene expression in heart tissue and some antioxidant indices in male rat. Sport Physiology. 2021;13(49):65-88. (In Persian).
  40. Nazem F, Hokkamian E, Ranjbar K, Nazari A. The effects of aerobic training on renal oxidative stress in myocardial infarction rats. Sport Physiology. 2017;9(34):79-94. (In Persian).
  41. Tolouei Azar J, Tofighi A, Arabzadeh E. The effect of 6 weeks endurance training on FSTL-1, NDNF, VEGF and vascular changes in healthy male rats. Sport Physiology. 2019;11(41):169-86. (In Persian).
  42. Yazdanian N, Asad MR, Rahimi M. The effect of high intensity interval training and moderate-intensity continuous training on PGC1Α and VEGF in heart muscle of male wistar rats. Sport Physiology. 2018;10(38):111-24. (In Persian).
  43. Ghasemi P, Gharakhanlo R, Molanouri Shamsi M, Khodadadi D. The effect of 4 weeks exercise on mRNA of GDNF in rat’s hippocampus with AD induced by Aβ1-42 injection. Sport Physiology. 2021;13(49):169-98. (In Persian).
  44. Omidi F, Hashemvarzi SA. The effect of eight weeks aerobic training with royal jelly consumption on some cardiovascular biomarkers during chronic high blood pressure induced by L-NAME in male rats. Sport Physiology. 2018; 9(36):143-58. (In Persian).
  45. Bahramvash Shams S, Farzanegi P, Azarbayjany MA. Effects of aerobic exercise and ethanolic extract of purslane seed on markers of oxidative stress and DNA damage in cardiac tissue of rats poisoned with hydrogen peroxide. Medical Laboratory Journal. 2021;15(3):40-6.

Pirooz M, Azarbayjani MA, Peeri M, Hosseini SA. The effect of aerobic training with vitamin d on extrinsic pathway of apoptosis and anti- apoptotic indices of heart tissue of rats exposed to oxidative damage induced by H2O2. J Report of Health Care. 2017;3(1):30-40. 

Sport Physiology
Volume 13, Issue 51 - Serial Number 51
November 2021
Pages 139-158

  • Receive Date 28 February 2021
  • Revise Date 28 August 2021
  • Accept Date 30 August 2021