Effect of Regular Aerobic Training and Garlic Extract Consumption on Plasma NO Levels and Tissue VEGF of the Soleus and Gastrocnemius Muscles in Elderly Rats

Authors
M. Najar-Shams
P. Farzanegi
Department of Exercise Physiology, Sari Branch, Islamic Azad University, Sari, Iran
Abstract
Objective: Elderly age is accompanied with reduced muscle strength and mass. Vascular endothelial growth factor (VEGF) is the most important stimulant of angiogenesis in muscles which, by increasing blood flow to the muscles, causes muscle strength and endurance. Exercise training leads to increased nitric oxide (NO) and release of VEGF by raising the shear stress during proliferation of endothelial cells and the process of angiogenesis. Garlic also modulates blood flow. This study has assessed the effects of regular swimming training combined with the consumption of garlic extract on the levels of NO plasma and VEGF tissue of soleus slow-twitch and gastrocnemius fast-twitch muscles in aged rats.
Methods: Male aged rats (40-50 weeks) with an average weight of 250-300 g were randomly divided into 5 groups (n=7 rats per group) - control, saline, aerobic training, garlic, and aerobic training + garlic. The swimming training program was scheduled for 50 m daily, 3 times per week for 8 weeks. The groups that received supplements and supplementary training were given a daily dose by gavage of 1 ml/kg body weight of the garlic extract for 8 weeks. We collected tissue and blood samples 48 h after the last training session and after a 10-12 h fasting period. The amount of NO and VEGF were detected by colorimetry and ELISA, respectively.
Results: One-way ANOVA revealed that regular aerobic training increased plasma NO (P=0.001), VEGF of the soleus muscle tissue (P<0.001), and VEGF of the gastrocnemius muscle (P=0.004) in the aged rats. In the group that received garlic extract, there was a significant increase in NO levels (P=0.001), and VEGF in the soleus muscle tissue (P=0.007) and gastrocnemius muscle tissue (P=0.015). Combined training and garlic supplement significantly increased the plasma level of NO (P<0.001), and VEGF in the soleus muscle tissue (P<0.001) and gastrocnemius muscle tissue (P=0.001) compared to the control and saline groups.
Conclusion: Consumption of garlic extract alone and combined with aerobic training significantly increased plasma NO levels, and VEGF of the soleus and gastrocnemius muscle tissues. However, while the extents of increase in the training combined with garlic extract group was higher than that of the garlic group, there was no significant difference observed between the two groups. The lack of significant difference between these two groups might be due to the intensity and type of the training and would need additional research.

Keywords

[1]  Rana SV. Metals and apoptosis: recent developments. J Trace Elem Med Biol 2008; 22(4): 262-84.
[2]  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.
[3]  Iversen N, Krustrup P, Rasmussen HN, Rasmussen UF, Saltin B, Pilegaard H. Mitochondrial biogenesis and angiogenesis in skeletal muscle of the elderly. Exp Gerontol 2011; 46(8): 670-8.
[4]  Hellsten Y, Rufener N, Nielsen JJ, Høier B, Krustrup P, Bangsbo J. Passive leg movement enhances interstitial VEGF protein, endothelial cell proliferation, and eNOS mRNA content in human skeletal muscle.Am J Physiol Regul Integr Comp Physiol 2008; 294(3): R975-82.
[5]  Hoier B, Prats C, Qvortrup K, Pilegaard H, Bangsbo J, Hellsten Y. Subcellular localization and mechanism of secretion of vascular endothelial growth factor in human skeletal muscle. FASEB J 2013; 27(9): 3496-504.
[6]  Kraus RM, Stallings HW 3rd, Yeager RC, Gavin TP. Circulating plasma VEGF response to exercise in sedentary and endurance-trained men. J Appl Physiol (1985) 2004; 96(4): 1445-50.
[7]  Egginton S. Invited review: activity-induced angiogenesis. Pflugers Arch 2009; 457(5): 963-77.
 [8]  Amaral SL, Sanchez LS, Chang AJ, Rossoni LV, Michelini LC. Time course of training-induced microcirculatory changes and of vegf expression in skeletal muscles of spontaneously hypertensive female rats. Braz J Med Biol Res 2008; 41(5): 424-31.
[9]  Tesauro M, Thompson WC, Rogliani P, Qi L, Chaudhary PP, Moss J. Intracellular processing of endothelial nitric oxide synthase isoforms associated with differences in severity of cardiopulmonary diseases: cleavage of proteins with aspartate vs. glutamate at position 298. Proc Natl Acad Sci U S A 2000; 97(6): 2832-5.
[10] Gentile C, dos Remedios C, Drake C, Davies M. The role of the VEGF/eNOS signaling pathway in cardiovascular development: a novel target to advance cardiovascular regeneration. FASEB J 2014; 28(1 Suppl): 1078.11.
[11] Ban CR, Twigg SM. Fibrosis in diabetes complications: pathogenic mechanisms and circulating and urinary markers. Vasc Health Risk Manag 2008; 4(3): 575-96.
[12] Schiller M, Javelaud D, Mauviel A. TGF-beta-induced SMAD signaling and gene regulation: consequences for extracellular matrix remodeling and wound healing. J Dermatol Sci 2004; 35(2): 83-92.
[13] Tang K, Xia FC, Wagner PD, Breen EC. Exercise-induced VEGF transcriptional activation in brain, lung and skeletal muscle. Respir Physiol Neurobiol 2010; 170(1): 16-22.
[14] Frisbee JC, Samora JB, Peterson J, Bryner R. Exercise training blunts microvascular rarefaction in the metabolic syndrome. Am J Physiol Heart Circ Physiol 2006; 291(5): H2483-92.
[15] Joyner MJ, Dietz NM. Nitric oxide and vasodilation in human limbs. J Appl Physiol (1985) 1997; 83(6): 1785-96.
[16] Seo DY, Lee S, Figueroa A, Kwak YS, Kim N, Rhee BD, Ko KS, Bang HS, Baek YH, Han J. Aged garlic extract enhances exercise-mediated improvement of metabolic parameters in high fat diet-induced obese rats. Nutr Res Pract 2012; 6(6): 513-9.
[17] Colín-González AL, Santana RA, Silva-Islas CA, Chánez-Cárdenas ME, Santamaría A, Maldonado PD. The antioxidant mechanisms underlying the aged garlic extract- and S-allylcysteine-induced protection. Oxid Med Cell Longev 2012; 2012: 907162.
[18] Al-Qattan KK, Thomson M, Al-Mutawa'a S, Al-Hajeri D, Drobiova H, Ali M. Nitric oxide mediates the blood-pressure lowering effect of garlic in the rat two-kidney, one-clip model of hypertension. J Nutr 2006; 136(3 Suppl): 774S-776S.
[19] Farzanegi P, Habibian M, Alinejad H. The Combined Effect of Regular Aerobic Exercise with Garlic Extract on Renal Apoptosis Regulatory Factors in Aged rats with Chronic Kidney Disease. Arak Medical University Journal 2016; 19(3): 62-70. (Persian)
[20] Slami AH, Habibian M, Farzanegi P. Effect of swimming exercise and garlic extract consumption on some of growth factors involved in angiogenesis and neurogenesis in the brain tissue of old rats. Daneshvar Med 2016; 23(121): 13-21. (Persian)
[21] Mounier R1, Pedersen BK, Plomgaard P. Muscle-specific expression of hypoxia-inducible factor in human skeletal muscle. Exp Physiol 2010; 95(8): 899-907.
[22] Peng CC, Chen KC, Hsieh CL, Peng RY. Swimming exercise prevents fibrogenesis in chronic kidney disease by inhibiting the myofibroblast transdifferentiation. PLoS One 2012; 7(6): e37388.
[23] Karimian J, Khazaei M, Shekarchizadeh P. Effect of Resistance Training on Capillary Density Around Slow and Fast Twitch Muscle Fibers in Diabetic and Normal Rats. Asian J Sports Med 2015; 6(4): e24040.
[24] Gavin TP, Drew JL, Kubik CJ, Pofahl WE, Hickner RC. Acute resistance exercise increases skeletal muscle angiogenic growth factor expression. Acta Physiol (Oxf) 2007; 191(2): 139-46.
[25] Higashi Y, Kihara Y, Noma K. Endothelial dysfunction and hypertension in aging. Hypertens Res 2012; 35(11): 1039-47.
[26] Jackaman C, Tomay F, Duong L, Abdol Razak NB, Pixley FJ, Metharom P, Nelson DJ. Aging and cancer: The role of macrophages and neutrophils. Ageing Res Rev 2017; 36: 105-16.
[27] Ziyadeh FN. Different roles for TGF-beta and VEGF in the pathogenesis of the cardinal features of diabetic nephropathy. Diabetes Res Clin Pract 2008; 82 Suppl 1: S38-41.
[28] Rodríguez I, González M. Physiological mechanisms of vascular response induced by shear stress and effect of exercise in systemic and placental circulation. Front Pharmacol 2014; 5: 209.
[29] Pearson T, Kabayo T, Ng R, Chamberlain J, McArdle A, Jackson MJ. Skeletal muscle contractions induce acute changes in cytosolic superoxide, but slower responses in mitochondrial superoxide and cellular hydrogen peroxide. PLoS One 2014; 9(5): e96378.
[30] Lieb W, Safa R, Benjamin EJ, Xanthakis V, Yin X, Sullivan LM, Larson MG, Smith HM, Vita JA, Mitchell GF, Sawyer DB, Vasan RS. Vascular endothelial growth factor, its soluble receptor, and hepatocyte growth factor: clinical and genetic correlates and association with vascular function. Eur Heart J 2009; 30(9): 1121-7.
[31] Dulak J, Józkowicz A, Dembinska-Kiec A, Guevara I, Zdzienicka A, Zmudzinska-Grochot D, Florek I, Wójtowicz A, Szuba A, Cooke JP. Nitric oxide induces the synthesis of vascular endothelial growth factor by rat vascular smooth muscle cells. Arterioscler Thromb Vasc Biol 2000; 20(3): 659-66.
[32] Wood RE, Sanderson BE, Askew CD, Walker PJ, Green S, Stewart IB. Effect of training on the response of plasma vascular endothelial growth factor to exercise in patients with peripheral arterial disease. Clin Sci (Lond) 2006; 111(6): 401-9.
[33] Olfert IM, Howlett RA, Tang K, Dalton ND, Gu Y, Peterson KL, Wagner PD, Breen EC. Muscle-specific VEGF deficiency greatly reduces exercise endurance in mice. J Physiol 2009; 587(Pt 8): 1755-67.
[34] van Weel V, Deckers MM, Grimbergen JM, van Leuven KJ, Lardenoye JH, Schlingemann RO, van Nieuw Amerongen GP, van Bockel JH, van Hinsbergh VW, Quax PH. Vascular endothelial growth factor overexpression in ischemic skeletal muscle enhances myoglobin expression in vivo. Circ Res 2004; 95(1): 58-66.
[35] Farzanegi P, Habibian M, Delavari H. The Effect of Aerobic Exercise on the Levels of Vascular Endothelial Growth Factor and Glucose in Hypertensive Postmenopausal Women: A Randomized Clinical Trial. Qom Univ Med Sci J 2014, 8(4): 6-12.(Persian)
[36] Fernandes T, de Castro Magalhães F, Crivoi do Carmo E, de Oliveira EM. Aerobic exercise training inhibits skeletal muscular apoptotic signaling mediated by VEGF-VEGR2 in spontaneously hypertesive rats. Rev Bras Med Esporte 2012; 18(6): 412-8.
[37] Hara Y, Noda A, Miyata S, Minoshima M, Sugiura M, Kojima J, Otake M, Furukawa M, Cheng XW, Nagata K, Murohara T. Effects of aged garlic extract on left ventricular diastolic function and fibrosis in a rat hypertension model. Exp Anim 2013; 62(4): 305-10.
[38] Saravanan G, Ponmurugan P. Ameliorative potential of S-allyl cysteine on oxidative stress in STZ induced diabetic rats. Chem Biol Interact 2011; 189(1-2): 100-6.
[39] Seo DY, Lee S, Figueroa A, Kwak YS, Kim N, Rhee BD, Ko KS, Bang HS, Baek YH, Han J. Aged garlic extract enhances exercise-mediated improvement of metabolic parameters in high fat diet-induced obese rats. Nutr Res Pract 2012; 6(6): 513-9.
[40] Takashima M, Kanamori Y, Kodera Y, Morihara N, Tamura K. Aged garlic extract exerts endothelium-dependent vasorelaxant effect on rat aorta by increasing nitric oxide production. Phytomedicine 2017; 24: 56-61.
[41] Dirsch VM, Kiemer AK, Wagner H, Vollmar AM. Effect of allicin and ajoene, two compounds of garlic, on inducible nitric oxide synthase. Atherosclerosis 1998; 139(2): 333-9.
Pathobiology Reserach
Volume 20, Issue 1
June 2017
Pages 63-76