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8 Weeks of 2S-Hesperidin Supplementation Improves Power Output at Estimated Functional Threshold Power and Maximum Power in Amat

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8 Weeks of 2S-Hesperidin Supplementation Improves Power Output at Estimated Functional Threshold Power and Maximum Power in Amat Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 16 September 2020 doi:10.20944/preprints202009.0341.v1 1 Article 2 8 Weeks of 2s-Hesperidin Supplementation Improves Power Output 3 at Estimated Functional Threshold Power and Maximum Power in 4 Amateur Cyclist 5 Full names of the authors and institutional/corporate affiliations 6 Francisco Javier Martínez Noguera *1, Cristian Marín Pagán 1, Jorge Carlos Vivas 7 2, Pedro E. Alcaraz 1. 8 1 Research Center for High Performance Sport. Catholic University of Murcia, campus de 9 los Jerónimos Nº 135 (UCAM, 30107, Murcia, Spain). 10 2 Health, Economy, Motricity and Education Research Group (HEME), Faculty of Sport 11 Sciences, University of Extremadura. Avda. de Elvas, s/n. (06006, Badajoz, Spain), 12 13 Contact details for the corresponding author 14 * Francisco Javier Martínez Noguera: Research Center for High Performance Sport. 15 Catholic University of Murcia, campus de los Jerónimos Nº 135 (UCAM, 30107, Murcia, 16 Spain); [email protected].; Tel.: +34 968 278 566 (F.J.M.N.) 17 18 Email addresses 19 FJMN: [email protected]. 20 CMP: [email protected]. 21 JCV: [email protected]. 22 PEA: [email protected]. 23 24 25 26 27 28 © 2020 by the author(s). Distributed under a Creative Commons CC BY license. Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 16 September 2020 doi:10.20944/preprints202009.0341.v1 29 ABSTRACT 30 2S-hesperidin is a flavanone (flavonoid) found in high concentrations in citrus 31 fruits. It has an antioxidant and anti-inflammatory effect, improving performance 32 in animals. This study investigated the effects of chronic intake of an orange extract 33 (2S-hesperidin) or placebo on aerobic-anaerobic and metabolic performance 34 markers in amateur cyclists. A double-blind, randomized, placebo-controlled trial 35 was carried out between late September and December 2018. Forty amateur 36 cyclists were randomized into two groups: one taking 500mg/day 2S-hesperidin 37 and other taking 500 mg/day placebo (microcellulose) for 8 weeks. All participants 38 completed the study. Performance and metabolic aerobic-anaerobic markers were 39 measured using incremental and rectangular tests by indirect calorimetry. The 40 anaerobic power was determined using Wingate tests. After 8 weeks 41 supplementation, there was a significant increase in the incremental test in 42 estimated functional threshold power (FTP) (3.23%; p≤0.05) and maximum power 43 (2.68%; p≤0.05) with 2S-Hesperdin compared to placebo. In the rectangular test, 44 there was a significant decrease in VO2 (-8.26%; p≤0.01) and VO2R (-8.88%; p≤0.01) 45 at VT2 in placebo; however, there were no significant differences between groups. 46 In the Wingate test, there was a significant increase (p≤0.05) in peak and relative 47 power in both groups, but without significant differences between groups. 48 Supplementation with an orange extract (2S-hesperdin) 500mg/day improves 49 estimated FTP and maximum power performance in amateur cyclists. 50 Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 16 September 2020 doi:10.20944/preprints202009.0341.v1 51 Keywords: flavonoid; polyphenols; orange extract; performance; endurance; 52 aerobic; anaerobic; nutrigenomic; sport nutrition 53 Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 16 September 2020 doi:10.20944/preprints202009.0341.v1 54 1. INTRODUCTION 55 Hesperidin is a flavonoid found mainly in citrus fruits [1], reaching high 56 concentration in sweet orange (Citrus sinensis) [2]. Due to its chemical structure, 57 including a chiral carbon (C-2), Hesperidin can be present as S or R isomer (Figure 58 1). 2S-hesperidin is the predominant natural form in citrus fruits [3], but industrial 59 processing leads to the transformation of the natural S isomer into the R isomer 60 (Figure 1) [4]. The bioavailability of the two isomers is different, for instance a 5.2- 61 fold higher efficiency in the glucuronidation has been observed for S-hesperetin 62 compared to R-hesperetin in vitro, without any significant change in the 63 sulfonation kinetics [5]. Clinical trials have demonstrated the therapeutic effects of 64 hesperidin and its metabolites in various diseases (e.g., neurological and 65 psychiatric disorders, cardiovascular diseases, etc.) due to its anti-inflammatory 66 properties, antioxidants, lipid reducers and insulin sensitizers [6-9]. In view of its 67 effects, the pharmaceutical and nutritional industries have extensively marketed 68 hesperidin. However, little attention has been paid to the effects of hesperidin on 69 physical performance. 70 **Insert figure 1** 71 72 Regarding performance, only one study has investigated the acute effect of 2S- 73 hesperidin in humans. Martínez et al. [10] showed that after ingesting one single 74 500 mg dose of either 2S-hesperidin or placebo (cross-over study) 5 hours before Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 16 September 2020 doi:10.20944/preprints202009.0341.v1 75 the test, trained cyclists significantly improved average power (2.3%), maximum 76 speed (3.2%) and total energy (∑ 4 sprint test) (2.6%) with Cardiose® 77 supplementation in the best sprint of the four repeated sprint test (30 s duration). 78 No significant changes were observed in any of these variables with placebo. 79 80 In humans, chronic supplementation of hesperidin has also been studied. Pittaluga 81 et al. [11] investigated the effect of 250 ml of red-orange juice (ROJ), which has a 82 high content of hesperidin, on exercise performance (incremental test) in healthy, 83 trained older women. Following 4 weeks of consumption of ROJ (3 per day), these 84 older women significantly increased their work capacity by 9.0% compared to 85 placebo (-1.5%). Another chronic study evaluated the effect of a 4-week 86 supplementation of 2S-hesperidin (500 mg/day) in trained cyclists and observed 87 significant increases in average power output (14.9 W = 5.0%) in a 10 min time-trial 88 test on a cycle ergometer, whereas those that consumed placebo had a non- 89 significant increase in average power output (3.8 W = 1.3%) [12]. 90 91 The effect of long-term intake of hesperidin has also been investigated in animal 92 studies. Biesemann et al. [13] observed that 6-weeks of hesperetin supplementation 93 (main metabolite of hesperidin) (50 mg·kg-1·d-1) improved running performance 94 (exercise time) in aged mice. De Oliveira et al. [14] found that four weeks of 95 hesperidin consumption (100 mg/kg body mass) enhanced the antioxidant 96 capacity in the continuous swimming group (183%) and decreased the lipid Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 16 September 2020 doi:10.20944/preprints202009.0341.v1 97 peroxidation (TBARS) in the interval swimming group (-45%) in rats. This study 98 also found an improvement in endogenous antioxidant enzymes, such as reduced 99 glutathione (GSH), oxidized glutathione (GSSG) and GSH:GSSG ratio. In the same 100 line, a recent study in trained animals reported that intake of hesperidin for 4 101 weeks improved performance and prevented immune alterations induced by 102 exhausting exercise [15]. Recently, one parallel-group study has shown 103 improvements in the time until exhaustion (58%) on maximal exercise test at 3 104 weeks of a 5-week chronic supplementation of 2S-hesperidin (200 mg/kg), but not 105 in placebo group [16]. In the same study, it was observed an enhancement of the 106 antioxidant state (superoxide dismutase (SOD), glutathione peroxidase (GPx)) in 107 the lymphoid and hepatic tissue after the test until exhaustion in the rats that 108 consumed 2S-hesperidin. 109 110 Another flavonoid, quercetin, has also demonstrated to improve the 5 km running 111 performance time (-11.3% quercetin group; -3.9% control group) after its 14 day 112 supplementation (250 mg/d) by trained triathletes [17]. A systematic review that 113 included 13 randomized controlled trials found that cocoa-derived flavonoid 114 (epicatechinand catechin, and oligomeric procyanidin) supplementation did not 115 affect performance [18]. Thus, there may be some specificity regarding the type of 116 flavonoid that affects physical performance. 117 118 The mechanisms by which chronic intake of hesperidin may improve performance Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 16 September 2020 doi:10.20944/preprints202009.0341.v1 119 are associated with increased activation of AMP-activated protein kinase (AMPK) 120 [19,20] and nuclear respiratory factor 2 (NRF2) [6], leading to improved 121 mitochondrial biogenesis and antioxidant status, respectively [21,22]. In addition, 122 hesperidin has the ability to improve nitric oxide synthesis (NO) [23], which may 123 improve glucose utilization in exercise and increase blood flow to the muscles, 124 promoting an increase in nutrient and oxygen delivery to the muscle [24]. More 125 detailed human studies are needed to determine precisely what molecular 126 mechanisms explain the effects of hesperidin. 127 128 It has been hypothesized that some molecules with anti-inflammatory and 129 antioxidant activity may interfere with exercise-generated adaptations causing a 130 decline in performance when ingested chronically [25]. However, there is 131 controversy on this issue, since supplementation of polyphenols, such as 132 quercetin, have been shown to improve performance [26]. To solve this question, 133 future studies on polyphenols (specifically flavonoids) are needed to clarify which 134 pathways or receptors are activated to help explain the possible or lack of 135 improvements in performance. 136 137 Based on the understanding behind the mechanism of hesperidin in vitro, as well 138 as the scientific evidence presented above, hesperidin is a good candidate for 139 improving performance. Hesperidin strongly increases intracellular ATP 140 compared to the AMPK activator 5-Aminoimidazole-4-carboxamide Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 16 September 2020 doi:10.20944/preprints202009.0341.v1 141 ribonucleotide (AICAR), even when AICAR concentration has been increased by 142 10-fold (100 µM) [13].
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