The Effect of Eight Weeks of High Intensity Interval Training with Citrus Aurantium Extract Consumption on Oxidative Stress and Antioxidant Levels of Soleus Muscle in Elderly Rats

Subject Areas : Open Access

Maryam Kheirdeh ¹ , Mehdi Noura ² , Rasoul Jamali Fashi ³ , Abdolali Rakhshanizadeh ⁴

1 - Physical Education Department of Sama Collage, Shiraz Branch, Islamic Azad University, Shiraz, Iran
2 - Department of Physical Education and Sport Sciences, Shiraz Branch, Islamic Azad University, Shiraz, Iran
3 - Department of Physical Education and Sport Sciences, Sama Technical and Vocational College, Varamin Branch, Islamic Azad University, Varamin, Iran
4 - Department of Physical Education and Sport Sciences, Payame Noor University, Iran

Received: 2020-11-29 Accepted : 2020-11-29 Published : 2020-11-21

Keywords: Antioxidant, Training, Oxidative stress, Aging, Citrus aurantium,

Abstract :

Abstract Introduction: Aging is a biological process that causes muscle dysfunction and muscle atrophy by increasing oxidative stress. Although the role of exercise and some herbs in improving muscle metabolism has been identified, the effect of high intensity interval training (HIIT) and Citrus aurantium (CA) consumption on oxidative stress in muscle tissue following aging is unknown. Therefore, the present study aimed to investigate the effect of eight weeks of HIIT with CA consumption on glutathione peroxidase (GPx), malondialdehyde (MDA) and carbonyl protein (PC) levels in the soleus muscle tissue of elderly rats. Methods: In this experimental study, 35 elderly rats with the age range of 14 to 18 months, and approximate weight of 270 to 320 g were randomly divided into (1) control (C), (2) sham (Sh), (3) HIIT, (4) CA and (5) HIIT+CA groups. Groups 3 and 5 performed HIIT for eight weeks, five sessions per week (at 85-110% VO2max and speed of 15-25 m/min). Also, groups 4 and 5 received 300 mg/kg/day Citrus aurantium extract peritoneally for eight weeks. To analyze the data, one-way analysis of variance with Tukey's post- hoc test was performed in Graph Pad Prism 8.3.0 software (P≥0.05). Results: HIIT increased GPx and decreased PC (P≥0.05). CA consumption and HIIT and CA interaction increased GPx and decreased MDA and PC in the soleus muscle tissue of elderly rats (P≥ 0.05). Conclusion: It seems that HIIT and CA consumption separately and synergistically have a beneficial effect on reducing oxidative stress and increasing antioxidant activity, however, further studies in this regard seem necessary. Keywords: Aging, Citrus Aurantium, Antioxidant, High Intensity Interval Training (HIIT), Muscle, Oxidative Stress   Extended Abstract Introduction Aging as a biological phenomenon causes muscle atrophy by decreasing mitochondrial function, increasing reactive oxygen/nitrogen species (RONS), decreasing superoxide dismutase (SOD), glutathione peroxidase (GPx), increasing malondialdehyde (MDA) and carbonyl protein (PC) (1). But regular exercise increases the expression of antioxidants in skeletal muscles by increasing mitochondrial biogenesis and protein kinases. High and moderate intensity training also increase SOD, GPx and PC. Aerobic, anaerobic and combined training increase MDA, GPx, SOD, and glutathione reductase (GR) (3, 7, 8) as well. Also, consumption of medicinal plants such as Citrus aurantium (CA) increases mitochondrial biogenesisand sirtuin 1. It reduces inflammatory factors, and has favorable effects on the elderly rats (9, 11). Due to the unknown effect of HIIT and CA consumption on antioxidant enzymes and oxidative stress in muscle tissue, the present study aimed to investigate the effect of eight weeks of HIIT and CA consumption on oxidative stress and antioxidant levels in the soleus muscle of elderly rats.   Methods In this experimental study, 35 elderly rats with the age range of 14 to 18 months, and approximate weight of 270 to 320 g were randomly assigned to (1) control (C), (2) sham (Sh), (3) HIIT, (4) CA and (5) HIIT + CA groups. Groups 3 and 5 performed HIIT for eight weeks, five sessions per week (at 85-110% VO2max and speed of 25-25 m/min) (12). Also, groups 4 and 5 received 300 mg/kg/day Citrus aurantium extract peritoneally for eight weeks (13). 48 hours after the last training session and by using a combination of ketamine and xylazine, the rats were anesthetized and their soleus muscles were extracted by the laboratory experts. To measure MDA levels, MDA measuring kit made by ZellBio GmbH, Ulm, with a sensitivity of 0.1 micromolar was used, and PC surfaces were measured by spectrophotometry using PC measuring kit made by Kiazist Iran company with a sensitivity of nanomoles per milligram. Also, GPx levels were measured using Nagpix ™ Glutathione Peroxidase kit using BMG Labtech Ltd., Aylesbury, UK. One-way analysis of variance with Tukey's post hoc test was used to analyze the results. Data analysis and plotting of research charts were performed using Graph Pad Prism 8.3.0 software (P≥0.05).   Results The mean and standard deviation of GPx, MDA and PC levels in the research groups are presented in Figures 1 to 3. The results of one-way analysis of variance showed a significant difference in GPx (P=0.001), MDA (P=0.001) and PC (P=0.001) levels in the muscle tissue of elderly rats in the research groups. The results of Tukey’s post- hoc test showed that GPx levels in the C and Sh groups were not significantly different (P=0.57), but in the HIIT (P=0.001), CA (P=0.001) and HIIT + CA (P=0.001) groups were significantly higher than the C group. In the HIIT (P=0.001), CA (P=0.001) and HIIT + CA (P=0.001) groups, the levels were significantly higher than the Sh group. No significant difference was observed in the CA and HIIT groups (P=0.99), but in the HIIT+CA group, the levels were significantly higher than the HIIT (P=0.001) and CA (P=0.001) groups (Figure 1). MDA levels in the C and Sh groups were not significantly different (P=0.99), but in the CA (P=0.02) and HIIT+CA (P=0.001) groups, they were significantly lower than the C group; also in the CA (P=0.007) and HIIT+CA (P=0.001) groups, they were significantly lower than the Sh group. MDA levels in the HIIT+CA group were significantly lower than the HIIT (P=0.001) and CA (P=0.003) groups (Figure 2).     PC levels in the C and Sh groups were not significantly different (P=0.23), however, in the HIIT (P=0.001), CA (P=0.001) and HIIT+CA (P=0.001) groups, they were significantly lower than the C and Sh groups. There was no significant difference between the CA (P=0.99) and HIIT+CA (P=0.91) groups compared to the HIIT group; Also, PC levels in the HIIT+CA group were not significantly different from the CA group (P=0.93) (Figure 3).   Discussion The results showed that HIIT increased GPx and decreased PC levels in the soleus muscle of elderly rats. HIIT with the mechanism of instant response to oxidative stress (OS) induced by exercise modulates cell redox increases protein kinases, the expression of respiratory transcription proteins (NRF1/2) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1), and modulates nuclear factor-κB (NF-κB). It also increases SOD and GPx expression and decreases PC and MDA (8, 14); Moreover, consumption of CA and HIIT + CA with the mechanism of increasing the expression of forkhead box O (FOXO) protein, activation of PI3K pathway, AKT1/2, IGF-1, increasing of PGC-1α, modulation of NRF1/2, NF-κB, increasing of protein synthesis and antioxidants enzymes in mitochondria and cytosol increases GPx and decreases MDA and PC in the soleus muscle tissue of elderly rats (9, 16).   Conclusion It seems that HIIT and CA consumption separately and synergistically have a beneficial effect on reducing oxidative stress and increasing antioxidant activity, however, further studies in this regard seem necessary.

References:

1. Kadoguchi T, Shimada K, Miyazaki T, Kitamura K, Kunimoto M, Aikawa T, et al. Promotion of oxidative stress is associated with mitochondrial dysfunction and muscle atrophy in aging mice. Geriatr Gerontol Int. 2020; 20 (1): 78– 84. [DOI:10.1111/ggi.13818] [PMID:31758637]


2. Sakellariou GK, Pearson T, Lightfoot AP, Nye GA, Wells N, Giakoumaki II, et al. Mitochondrial ROS regulate oxidative damage and mitophagy but not age-related muscle fiber atrophy. Sci Rep. 2016; 6: 33944. [DOI:10.1038/srep33944] [PMID:27681159] [PMCID:PMC5041117]


3. Gorni D, Finco A. Oxidative stress in elderly population: A prevention screening study. Aging Med. 2020; 3 (3): 205– 213. [DOI:10.1002/agm2.12121] [PMID:33103041] [PMCID:PMC7574639]


4. Bernardi M, Peluso I. Interactions between oxidative stress and cardiorespiratory fitness: old and new biomarkers. Curr Opin Toxicol. 2020; (20-21): 15- 22. [DOIorg/10.1016/j.cotox.2020.03.005]


5. Yavari A, Javadi M, Mirmiran P, Bahadoran Z. Exercise-induced oxidative stress and dietary antioxidants. Asian J Sports Med. 2015; 6 (1): e24898. [DOI:10.5812/asjsm.24898][PMID:25883776] [PMCID:PMC4393546]


6. Hsieh S-S, Chueh T-Y, Huang C-J, Kao S-C, Hillman CH, Chang Y-K, et al. Systematic review of the acute and chronic effects of high-intensity interval training on executive function across the lifespan. J Sports Sci. 2020; 1– 13. [DOI:10.1080/02640414.2020.1803630] [PMID:32780634]


7. Paes L, Lima D, Matsuura C, De Souza MDG, Cyrino F, Barbosa C, et al. Effects of moderate and high intensity isocaloric aerobic training upon microvascular reactivity and myocardial oxidative stress in rats. PLoS One. 2020; 15 (2): e0218228. [DOI:10.1371/journal.pone.0218228] [PMID:32032358] [PMCID:PMC7006926]


8. Ammar A, Trabelsi K, Boukhris O, Glenn JM, Bott N, Masmoudi L, et al. Effects of aerobic-, anaerobic-and combined-based exercises on plasma oxidative stress biomarkers in healthy untrained young adults. Int J Environ Res Public Health. 2020; 17 (7): 2601. [DOI:10.3390/ijerph17072601] [PMID:32290148] [PMCID:PMC7178085]


9. Shykholeslami Z, Abdi A, Barari A, Hosseini SA. The effect of aerobic training with Citrus aurantium L. on SIRT1 and PGC-1α gene expression levels in the liver tissue of elderly rats. Jorjani Biomed J. 2019; 7 (4): 57– 65. http://goums.ac.ir/jorjanijournal/article-1-720-en.html


10. Elyasi L, Ghazvini H. The protective effects of Citrus aurantium extract on a 6-hydroxydopamine- induced model of parkinson’s disease in male rats. Anat Sci J. 2020; 17 (1): 1– 6. http://anatomyjournal.ir/article-1-230-en.html


11. Testai L, Piragine E, Piano I, Flori L, Da Pozzo E, Miragliotta V, et al. The Citrus flavonoid naringenin protects the myocardium from ageing- dependent dysfunction: potential role of SIRT1. Oxid Med Cell Longev. 2020; 1- 15. [DOI:10.1155/2020/4650207]


12. Yazdanparast Chaharmahali B, Azarbayjani MA, Peeri M, Farzanegi Arkhazloo P. The effect of moderate and high intensity interval trainings on Cardiac apoptosis in the old female rats. Rep Heal Care. 2018; 4 (1): 26– 35. http://jrhc.miau.ac.ir/article_2873.html


13. He W, Li Y, Liu M, Yu H, Chen Q, Chen Y, et al. Citrus aurantium L. and its flavonoids regulate TNBS-induced inflammatory bowel disease through anti-inflammation and suppressing isolated jejunum contraction. Int J Mol Sci. 2018; 19 (10): 3057. [DOI:10.3390/ijms19103057] [PMID:30301267] [PMCID:PMC6213068]


14. Davari F, Alimanesh Z, Alimanesh Z, Salehi O, Hosseini SA. Effect of training and crocin supplementation on mitochondrial biogenesis and redox-sensitive transcription factors in liver tissue of type 2 diabetic rats. Arch Physiol Biochem. 2020; 1– 6. [DOI:10.1080/13813455.2020.1762663] [PMID:32401063]


15. Fakhri S, Shakeryan S, Alizadeh A, Shahryari A. Effect of 6 weeks of high intensity interval training with nano curcumin supplement on antioxidant defense and lipid peroxidation in overweight girls- clinical trial. Iran J diabetes Obes. 2020; 1- 10. [DOI:10.18502/ijdo.v11i3.2606]


16. Zhu Q, Qu Y, Zhou X-G, Chen J-N, Luo H-R, Wu G-S. A dihydroflavonoid naringin extends the lifespan of C. elegans and delays the progression of aging- related diseases in PD/AD models via DAF-16. Oxid Med Cell Longev. 2020; 6069354. [DOI:10.1155/2020/6069354] [PMID:32832002] [PMCID:PMC7422489]


17. Hosseini SA, Hamzavi K, Safarzadeh H, Salehi O. Interactive effect of swimming training and fenugreek (Trigonella foenum graecum L.) extract on glycemic indices and lipid profile in diabetic rats. Arch Physiol Biochem. 2020; 1– 5. [DOI:10.1080/13813455.2020.1826529] [PMID:33017260]


18. Ana L, Lima MJ, Simoni R. Heart rate variability in the frequency domain after strength training with citrus auratium supplementation. Int Phys Med Rehab J. 2020;5(3):110–113. [DOI:10.15406/ipmrj.2020.05.00240]


19. Hansen DK, George NI, White GE, Abdel-Rahman A, Pellicore LS, Fabricant D. Cardiovascular toxicity of Citrus aurantium in exercised rats. Cardiovasc Toxicol. 2013;13(3):208–219. [DOI:10.1007/s12012-013-9199-x] [PMID:23397375]