Effects of Chronic Beetroot Juice Supplementation on Maximum Oxygen Uptake, Velocity Associated with Maximum Oxygen Uptake
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European Journal of Applied Physiology (2019) 119:1043–1053 https://doi.org/10.1007/s00421-019-04094-w ORIGINAL ARTICLE Effects of chronic beetroot juice supplementation on maximum oxygen uptake, velocity associated with maximum oxygen uptake, and peak velocity in recreational runners: a double-blinded, randomized and crossover study Talitha F. de Castro1 · Francisco de Assis Manoel2 · Diogo H. Figueiredo2 · Diego H. Figueiredo2 · Fabiana Andrade Machado1,2 Received: 1 August 2018 / Accepted: 6 February 2019 / Published online: 12 February 2019 © Springer-Verlag GmbH Germany, part of Springer Nature 2019 Abstract Purpose This study investigated the effects of chronic 3-day beetroot juice (BRJ) supplementation on maximum oxygen uptake (VO2max), velocity associated with VO2max (vVO2max), and peak velocity (Vpeak) in recreational runners. Methods Thirteen male recreational runners (age 28.2 ± 3.0 years, height 176.8 ± 0.1 cm, body mass 74.4 ± 9.5 kg) performed four tests on a treadmill in a randomized, double-blind, crossover design: two maximum incremental tests to determine VO2max − and vVO2max, and two tests to determine Vpeak. Trials were performed following 3 days of supplementation of NO 3 -rich BRJ − − 1 − − − 1 in natura (8.4 mmol NO3 day ) or BRJ NO3 -depleted placebo (0.01 mmol NO3 day ), with the last dose being ingested 2 h before each test. During the tests, maximum heart rate (HR max), maximal rating of perceived exertion (RPE max), pre- and post-test glucose concentrations (Glucpre, Glucpost), and peak blood lactate concentration were determined. − 1 − 1 Results VO2max was higher following BRJ vs PLA (46.6 ± 6.4 vs 45.1 ± 5.8 mL kg min ; P = 0.022), as well as vVO2max − 1 − 1 (14.5 ± 0.8 vs 13.9 ± 1.0 km h P = 0.024) and Vpeak (15.5 ± 1.1 vs 15.2 ± 1.2 km h P = 0.038), with no differences in the other variables. − − − 1 Conclusion Consumption of NO 3 -rich BRJ in natura (8.4 mmol NO 3 day ) once per day for 3 days improved VO2max, vVO2max and Vpeak in recreational runners without changing the other analyzed variables. Keywords Nitrate supplementation · Nitric oxide · Running · Exercise nutritional science · Physical endurance Abbreviations ES Effect size LA Blood lactate concentration Glucpre Pre-glucose concentrations LApeak Peak blood lactate concentration Glucpost Post-glucose concentrations ATP Adenosine triphosphate HR Heart rate ADP Adenosine diphosphate HRmax Maximum heart rate BRJ Beetroot juice supplementation Inc Speed increment − NaNO3 Sodium nitrate NO Nitric oxide Communicated by Anni Vanhatalo. NOS Nitric oxide synthase − * Fabiana Andrade Machado NO2 Nitrite − [email protected] NO3 Inorganic nitrate 1 O2 Oxygen Post-graduate Program of Physiological Sciences, PCr Phosphocreatine Department of Physiological Sciences, State University of Maringá-PR, Colombo Avenue, 5790, Maringá, Pi Inorganic phosphate PR 87020-900, Brazil PLA Placebo 2 Associate Post-graduate Program in Physical Education P/O Phosphate/oxygen ratio UEM/UEL, Department of Physical Education, State RE Running economy University of Maringá-PR, Colombo Avenue, 5790, Maringá, RPE Rating of perceived exertion PR 87020-900, Brazil Vol.:(0123456789)1 3 1044 European Journal of Applied Physiology (2019) 119:1043–1053 RPEmax Maximal rating of perceived exertion C oxidase (Brown and Cooper 1994; Cleeter et al. 1994), SIRT1 Regulatory protein sirtuin resulting in “local hypoxia” (Larsen et al. 2011) and con- T Number of seconds required to complete a sequent increase in tissue availability (Tong et al. 2011). stage Regarding the improvement of muscular function, besides its t Number of seconds sustained during the incom- role in the modulation of the contractile function, especially plete stage in type II muscle fibers (Ferguson et al. 2013; Hernandés Vcomplete Running velocity of the last complete stage et al. 2012), it is believed that NO can match blood flow VO2 Oxygen uptake with local demand for O 2 providing the best distribution VO2max Maximum oxygen uptake (Hord et al. 2009). Vpeak Peak velocity It is believed that in addition to directly impacting O2 vVO2max Velocity associated with VO2max cost of endurance running at a given submaximal speed, − NO3 supplementation may improve other physiologi- cal determinants of endurance exercise such as maximal Introduction oxygen consumption (VO2max) (Joyner and Coyle 2008; Dominguez et al. 2017; Vanhatalo et al. 2011). This variable Beetroot juice (BRJ) supplementation has been widely is frequently used to indicate the cardiorespiratory fitness of − studied after evidence showed that inorganic nitrate (NO 3 ) an individual because it is quantitatively and qualitatively − consumption increased plasma concentrations of NO3 and related to metabolic and cardiovascular changes (Edvard- − nitrite (NO2 ) in a dose-dependent manner (Nyakayiru et al. sen et al. 2014; Bassett and Howley 2000). An individual’s 2017; Wylie et al. 2013). This supplementation could also VO2max is determined by genetic factors, age, gender, and induce the production of nitric oxide (NO) independent of level of training (Rowland 1996). Among its limiting factors the conventional L-arginine pathway. This occurs mainly are pulmonary ventilation, alveolar–capillary O2 diffusion, under conditions of hypoxia and low pH, when the activity cardiovascular system, and the arteriovenous O 2 difference of the nitric oxide synthase (NOS) enzyme family, depend- (Saltin and Strange 1992). It is directly affected by vasodi- ent on oxygen (O2), is impaired (Modin et al. 2001; Moncada lation and muscular vasoconstriction, and also the capacity and Higgs 1993). NO is an important signaling molecule of energy generation by oxidative phosphorylation (Amann that promotes relaxation of vascular smooth muscle and 2012; Denadai 1999; Holloszy and Coyle 1984). In addition, increases tissue blood flow mediated by guanylyl cyclase VO2max is considered a useful variable to predict endurance (Francis et al. 2010; Vanni et al. 2007). In addition to its performance (Midgley et al. 2007; Billat et al. 1999). direct effect on vasodilation and hemodynamics (Govoni From the determination of VO2max it is possible to iden- et al. 2008), NO plays a key role in many physiological pro- tify other variables such as the velocity of VO2max occur- cesses that may impact endurance exercise performance and rence (vVO2max), which in physiological terms reflects the even facilitates adaptation to exercise training (de Castro interaction between endurance running at a given submaxi- et al. 2018; Thompson et al. 2017; Bailey et al. 2009). In mal speed and VO2max (Buchheit al. 2010; Almarwaey et al. − addition, chronic and acute dietary NO 3 supplementation 2004; Billat and Koralsztein 1996). Moreover, vVO2max can reduces the O 2 cost of exercise and increases exercise effi- explain variations in performance that endurance running at ciency, as is reported in many (Whitfield et al. 2016; Lansley a given submaximal speed and VO2max alone would not be et al. 2011; Larsen et al. 2007, 2011; Bailey et al. 2009, able to do (Hill and Rowell 1996). 2010; Vanhatalo et al. 2010), although not in all studies Another variable that has gained attention among (Balsalobre-Fernández et al. 2018; Betteridge et al. 2016; researchers, trainers, and endurance runners is the maxi- Thompson et al. 2015). mal velocity attained during an incremental test (Vpeak) Some mechanisms are proposed to explain this ergogenic (McLaughlin et al. 2010; Saunders et al. 2010). Previ- effect on endurance performance and on the cardiorespira- ous studies have shown that Vpeak reflects the intensity of tory components of aerobic exercise: reduction of the cost VO2max and vVO2max; however, it is determinate and not esti- of ATP production of contractile force and attenuation of mated, thus associating itself directly with aerobic power the stimulus for oxidative phosphorylation and oxygen (da Silva et al. 2015). In addition, Vpeak is a great predic- uptake (VO2) (Vanhatalo et al. 2011; Bailey et al. 2010), tor of endurance performance over 3–90 km (Machado an increase in the mitochondrial phosphorylation efficiency, et al. 2013; Midgley et al. 2007; Mclaughlin et al. 2010), evaluated by the phosphate/oxygen ratio (P/O) ratio (Bailey and like vVO2max, is considered a good variable to moni- − et al. 2009). In addition, there is evidence that NO 2 can tor and prescribe endurance training (Manoel et al. 2017, act as an alternative acceptor of electrons, thus replacing 2018; Da Silva et al. 2017). Since determination of Vpeak the role of O 2 in respiration (Bailey et al. 2009; Basu et al. does not require ergospirometry, it is considered a very 2008; Brown 2001) and inhibits O 2 binding at cytochrome practical and attractive variable compared to vVO2max. In 1 3 European Journal of Applied Physiology (2019) 119:1043–1053 1045 − addition, different from the determination of vVO2max, which vasodilatory properties of NO 3 and their influence onO 2 considers the minimum velocity that VO2max reached dur- use during skeletal muscle contraction, and the mitochon- ing an incremental test to exhaustion, independent of the drial benefits and their relationship with the performance time remained in the stage that it occurs (Billat et al. 1994, variables, we hypothesized that after chronic BRJ supple- 1996), the determination of Vpeak considers every second mentation VO2max, vVO2max and Vpeak would be increased remained during the test, even if the last stage is not com- compared to PLA condition. pleted adjusted with Kuipers et al. (2003) equation, which in turn may be more sensitive to detect the effects of train- ing. For a proper training prescription, it is necessary to Methods use variables that control and monitor the intensity of effort and possible physiological adaptations resulting from this Participants and design practice and, most importantly, that show a correlation with performance. Thus, vVO2max and Vpeak are the preferred vari- Thirteen male recreational runners (age 28.2 ± 3.0 years, ables to monitor and to prescribe training among endurance height 176.8 ± 0.1 cm, body mass 74.4 ± 9.5 kg) with 10-km athletes.