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© MEDSPORTPRESS, 2016; 2(2); Vol. 7, 103-108 DOI: 10.5604/20815735.1232424

ORIGINAL ARTICLE

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Effects of a training session on IGF-I, IGFBP3, CK and LDH of brazilian jiu-jitsu fighters Hugo Tourinho Filho1(A,B,C,D), Enrico F. Puggina1-2 (A,B,D), Camila de Moraes1 (C,D), Lucas de P. Oliveira2 (A), Marcela Pires2 (A,B), Carlos Eduardo Martinelli Júnior2(A,C) 1 2

School of Physical Education and Sport of Ribeirao Preto, University of Sao Paulo, Ribeirao Preto, SP, Brazil Ribeirao Preto Medical School, University of Sao Paulo, Ribeirao Preto, SP, Brazil

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Key words: endocrinology, training and brazilian jiu-jitsu

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Summary

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Introduction. Exercise is related to anabolism induced by the GH/IGF-I axis, which is responsive to high intensity training. However, the literature is ambiguous regarding the behavior of the GH/IGF-I axis during exercise. While some studies shows increased levels of these hormones, other studies have found reduced levels of IGF-I after training sessions. From these statements, the objectives of this study were to investigate the effects of a Jiu-Jitsu training session on the IGF-I, IGFBP-3, CK and LDH concentrations. Material and methods. Nine male jiu-jitsu fighters participated in the study, they represent the national elite of the sport in Brazil, with 5.4±2.7 years of practice. Blood samples were collected in the beginning and at the end of a training session. IGF-I, IGFBP3, CK and LDH were assayed by colorimetric methods. Results. were analyzed using Wilcoxon test (p< 0.05). Results: No difference was observed regarding IGF-I (p= 0.57) and IGFBP-3 (p= 0.73) levels before and after the Jiu-Jitsu training session. A jiu-jitsu training session composed by six fights of 7.5 minutes with perception of effort between “hard” and “very hard” did not change the IGF- I and IGFBP-3 concentrations. Regarding CK activity, increases were observed (p= 0.002) when comparing the situations before and after training session, while LDH enzyme activity was not affected (p= 0.80). Conclusions. The absence of reduced values of IGF-I and IGFBP-3 in the second measurement would be explained by the training status of the athletes, who were in their maximum level of performance in the season.

Introduction

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Exercise is closely related to anabolic functions caused by the action of the GH / IGF-I axis, that is characterized as a system of growth mediators, receptors and binding proteins that control somatic and tissue growth in many species. Basal levels of IGF-I are positively correlated with muscle mass and physical fitness in children, adolescents and adults (ELIAKIN & NEMET, 2013; Eliakin, Nemet, & Cooper, 2005; Martinelli JR, Custódio, & Aguiar-Oliveira, 2008; Poehlman & Copeland, 1990; Tirakitsoontorn, Nussbaum, Moser, Hill, & Cooper, 2001). In the last years, the literature has been ambiguous regarding the behavior of the GH/IGF-I axis during exercise. Some studies shows increased levels of these hormones, specifically the GH levels (A. Eliakim et al., 2009; Mejri, Bchir, Ben Rayana, Ben Hamida, & Ben Slama, 2005; Nemet et al., 2012) while other studies have found reduced levels of IGF-I after training sessions (A. Eliakim, Brasel, Mohan,

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Journal of Combat Sports and Martial Arts

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Wong, & Cooper, 1998; A. Eliakim, Scheett, Newcomb, Mohan, & Cooper, 2001; Pilz-Burstein et al., 2010). Few studies investigated the hormonal responses associated to the effects of combat sports practice in this variables (Pilz-Burstein et al., 2010). In the specific case of Brazilian Jiu-Jitsu, it is clear that this sport has gained prominence in Brazil and increased worldwide interest. However, most of the physical and technical trainers do not know the physiological mechanisms manifested during the combat, which predispose them to train their athletes empirically, based on knowledge brought and perpetuated for decades (Del Vechio, Bianchi, Hirata, & Chacon-Mikahil, 2007). Considering the relationship between load and intensity of exercise, training status and changes in GH/IGF-I action, the assessment of possible changes in IGF-I, IGFBP3, CK and LDH concentrations could be interesting as these variables can directly affect the performance of athletes. (EliakimM & Nemet, 2010; Elloumi et al., 2005). The status of the GH/IGF-I axis may reflect training overload condition and it can be considered as a sen-

n=9 25 ± 4.7 71.3 ± 8.0 176.0 ± 5.0 5.4 ± 2.7

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Variables Age (years) Weight (kg) Height (cm) Practice Experience (years)

Imunoassays

sitive marker for excessive training load beside providing information about the nutritional status of the fighter (A. ELIAKIM & NEMET, 2012). It is particularly important in combat sports where intense training is combined with caloric restriction. This prompted us to investigate the effects of Brazilian Jiu-Jitsu training session on the IGF-I, IGFBP-3, CK and LDH concentrations.

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Determination of serum IGF-I and IGFBP3 Serum IGF-I and IGFBP-3 concentrations were measured by a solid-phase, enzyme-labled chemiluminescent immunometric assay (Immulite 2000, Siemens). IGFBPs interferences were avoided by acid-ethanol extraction. All samples were determined in the same assay. The intra-assay coefficient of variation was 3.0 and 4.5 % for IGF-I and IGFBP-3 assays, respectively. The sensitivity of the assays were 20 ng/ml and 0.1 mg/l, respectively. The tests were carried out in the Laboratory of Pediatrics of the Clinical Hospital of Medical School of Ribeirao Preto, University of São Paulo.

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Determination of plasmatic ck and ldh activity Plasmatic activities of CK and LDH were determined using commercial kits from Bioliquid® (Pinhais, Brasil). The assays were carried following the methods proposed by the manufacturer and read at 340 nm wavelength and 27ºC (spectrophotometer Ultrospec 3100 Pro from Amersham Pharmacia Biotech®, Piscataway, EUA). All samples were determined in the same assay. The intra-assay coefficient of variation was 2.5 and 1.5 % for CK and LDH assays, respectively..

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Study design Non-fast blood samples were collected from the subjects before the training session, preceded by 30 minutes of rest, and a new sample was obtained immediately after the training session.

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Material and methods Subjects A convenience sample of 9 male adult Brazilian Jiu-Jitsu fighters with regular training frequency were non-randomly selected. The sample size was determined the availability and acceptance to participate in the study. Written consent was obtained from all participants. The protocol was approved by the local ethics committee in accordance with the Helsinki Declaration. Subject characteristics are shown in Tab. 1.

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Tab. 1. Clinical characterization of the 9 Jiu-Jitsu fighters. Data are expressed as mean ± SD.

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Brazilian Jiu-Jitsu Training Session The training session used in this study was composed by a warm up (20 minutes of stretching exercises); a main part divided between the improvement of techniques (20min) and then a sequence composed by six Brazilian Jiu-Jitsu fights, with a duration of 7.5 minutes per fight, totaling 45 minutes of effort. At the end of the training, the athletes did stretching exercises. The athletes reported the training intensity using Borg scale for perceived exertion (Borg, 1982).

Blood collection Venous blood collection was obtained by venipuncture. The blood samples collection was carried in a room located at the fighters training center. Five mililiters of blood were collected from each athlete in tubes with and without anticoagulant (EDTA) and were immediately stored in ice (between 0-4°C) and transported to the laboratory to be centrifuged. After centrifugation, serum and plasma were obtained and stored at -80 ° C for futher analysis.

24H diet recall Multiple surveys were administered aiming to quantify the amount of food consumed in 24 hour period prior to the interview (Marchioni, Slater, & Fisberg, 2004). Six surveys was obtained from each fighter during the week before the Brazilian Jiu-Jitsu Championship. To calculate the data obtained from multiple 24 hours dietary recalls, the DietWin® program was used, which is a support system for nutritional assessment and diet prescription. The parameters established by the American Dietetic Association and Dietary Reference Intakes (DRIs) for age and level of physical activity performed were used to analyze the data obtained (Marchioni et al., 2004; P, 2002 )

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Statistics In order to compare the acute effects of a Brazilian Jiu-Jitsu training session on circulating IGF-I levels, IGFBP3, LDH and CK two statistical methods were used. Non-parametric Wilcoxon test was used for IGF-I and IGFBP3 data, while paired t-test was used for CK and LDH. The results were expressed as mean and standard deviation (SD). Statistical differences were assumed for p < 0.05.

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Filho H. T. et al. IGF-I, IGFBP3, CK and LDH in fighters

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Fig. 3. CK activity before and after Brazilian Jiu-Jitsu training session. n = 9. * p = 0.002

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Fig. 2. IGFBP3 serum concentrations before and after a Brazilian Jiu-Jitsu training session. n = 9. p = 0.73

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Fig. 4. LDH activity before and after Brazilian Jiu-Jitsu training session. n = 9. p = 0.80

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Fig. 1. IGF-I serum concentration before and after a Brazilian Jiu-Jitsu training session. n = 9. p = 0.57

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141.9 ± 30.1; female - 64.1 ± 30.6 mcg / dL) in both sexes. The fight simulation protocol also significantly reduced testosterone levels (men - 1.9 ± 1.6; female – 0.02 ± 0.06 ng / mL). In summary the authors concluded that the stimuli from the protocol developed with Taekwondo fights led to a typical hormonal response of a catabolic state. They also state that such hormonal responses can affect not only the muscular system as other systems (immune, cardiorespiratory) and therefore the hormonal responses can be used as a complementary tool to enhance the sports training cycles and monitor the training load. Steinacker, Reissnecker and Liu (2004) additionally say that understanding the function of hormones, cytokines and regulatory function that muscular system can perform during exercise practice would be critical in obtaining a deeper understanding of the complex adaptation mechanism of the body to exercise and physical training. Current literature has been signaling to a number of factors that can influence the behavior of the GH / IGF-I axis beyond exercise, standing out the reduction of pre competition weight (Nmet, Pntello, Rose-Groton, & Cooper, 2004), high carbohydrate and lipids diet pre-training (Galassetti et al., 2006), overtraining (Nmet et al., 2004) and pituitary refraction (A Eliakim, Brasel, & Cooper, 1999). The analysis of the dietary recall performed with athletes before the Brazilian n Jiu-Jitsu Championship allows to exclude the influence of the diet rich in carbohydrates and lipids, however, the weight reduction and overtraining appear as variables that can attenuate the IGF-I levels post-training. The training session used in this study is quite common among Jiu-Jitsu figthers and was classified as hard and very hard by the athletes (Borg, 1982). Regarding the athletes diet, it was possible to identify that all fighters revealed reduced caloric intake in relation to the minimum recommended for sex, age and fitness according to the parameters established by the guidelines of the American Dietetic Association (P, 2002 ). It is important to note that even in intense training conditions and under reduced caloric intake, a reduction in IGF-I levels and IGFBP-3 was not observed at the end of the training session. A possible hypothesis to explain the absence of suppression of the GH/IGF-I axis verified in this study is the athletes training status. It is possible to argue that in our study, the fighters were in their maximum level of fitness in the season, since they were near the main competition (Brazilian Jiu-Jitsu championship aiming the classification to the World Championship). In this stage, it is expected that the athletes are in high state of training as a result of a tapering strategy performed during the systematization of training (Maglischo, 2010). Although it is not an objective of the study, it is important to note that the next week after the data collection of the study, three of nine subjects of the sample won the Brazilian championship and one of them became also a world champion. In addition, all other participants performed very well in the championships, losing fights for only those who later would be the champions.

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Results

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Training intensity Using Borg scale for perceived exertion (PSE), the athletes reported the training intensity between 15 to 17, Hard and Very Hard (Borg, 1982).

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IGF-I AND IGFBP-3 No difference was observed in IGF-I concentrations when rest (before training) and after training session samples were compared (p = 0.57; Fig. 1). No difference was also found regarding IGFBP-3 concentrations when rest and after training session samples were compared (p = 0.73; Fig. 2).

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CK AND LDH With respect to CK enzyme activity levels, significant increases were observed (p = 0.002) when comparing the situations before and after training session (Fig. 3), while LDH enzyme activity did not show significant increases (p = 0.80) in its quantification (Fig. 4).

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In our study, it was not possible to observe changes in the behavior of IGF-I and IGFBP3 binding protein before and after a training session held in the week before the Brazilian Jiu-Jitsu championship. Contrary to the data of this study, (Nemet, Oh, Kim, Hill, & Cooper, 2002) observed the effects of a typical training session of wrestling on the GH / IGF-I axis in adolescents and found significant decreases in levels of the following anabolic mediators: total IGF-I (-11.2 ± 2, 3%), IGF-I bound (-11.2 ± 2.4%) and insulin (-42 ± 10%). Notably, increases were also observed in the following cytokines; IL-6 (795 ± 156%), TNF (30 ± 12%), IL-1β (286 ± 129%), and increased levels of IGFBP-1 binding protein, known as an inhibitor of IGF-1 effects (Frystyk, 2010). To Nemet et al. (2002), these data demonstrate that an intense exercise bout in wrestling young athletes led to a reduction in the levels of anabolic mediators and a marked increase in cytokines, leading to a predominantly catabolic response. Corroborating the data brought by Nemet et al. (Nemet et al., 2002), Pilz-Burstein et al. (Pilz-Burstein et al., 2010) evaluated the effects of a Taekwondo fight simulation (three fights of 6 minutes with a 30 minutes interval between the fights) on anabolic hormones IGF-I, LH, FSH, estradiol and testosterone and the catabolic hormone cortisol in adolescents fighters from national elite (aged 12 to 17 years, n = 10 boys and 10 girls). The authors found that the combat simulations led to a significant decrease (p < 0.05) in IGF-I levels (male - 27.1 ± 25.6; female - 22.4 ± 36,3ng / ml) LH (male - 0.7 ± 1.2; female - 2.3 ± 3.3 U / L), and FSH (male - 0.9 ± 0.5; female - 1.5 ± 1.1 U / L), and a significant increase (p < 0.05) in the cortisol levels (male -

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Discussion

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arbitrated in accordance with the rules of Brazilian Jiu-Jitsu, was possible to identify significant increases in the concentrations of CK and LDH at the end of two consecutive matches of 10 minutes, compared to the pre-competition values (Coswig, Neves, & Del Vecchio, 2013). Although there is disagreement in the literature regarding indirect markers of muscle damage would rise after competitions or training sessions in Jiu-Jitsu, there seems to be consensus that the fighting results in muscle damage (Andreato et al., 2013; Coswig et al., 2013; Kasperska, Żurek, & Zembron-Łacny, 2011). Muscle damage can be a result of changes in the membrane, hypoxia and ischemia of the muscle cell resulting from isometric and dynamic actions that are developed during the sport practice. Coswig, Neves and Del Vecchio (Coswig et al., 2013) also point out that this muscle damage may be associated with anti-inflammatory signaling and increased immunological actions.

Conclusions

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An acute session of Brazilian Jiu-Jitsu (BJJ) training, consisting in the main part of six fights with 7.5min total duration with perceived exertion between “hard” and “Very Hard” resulted in significant elevations in plasma CK activity, but did not change the behavior of LDH, IGF-I and its IGFBP3 hormone binding protein. A possible explanation for the absence of reduced values of IGF-I and IGFBP-3 in the second measurement in this study would be the training status of the Jiu-Jitsu fighters, who were in their maximum level of performance in the season. The responses observed in the GH/ IGF-I axis components can be used as training status markers in fighting sports.

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In his sense, these findings demonstrate that the behavior of the components of the GH / IGF-I axis may be used as an important marker of training status. Reinforcing this idea Eliakim and Nemet (ELIAKIN & NEMET, 2013) state that the collection of baseline and changes in the GH / IGF-I axis behavior when compared to the results obtained in past seasons of training, combined with the knowledge and experience of past experiences can serve as a important tool to be used by coaches as load control parameters during the training season. In the present study increases in CK activity were found after an acute Brazilian Jiu-Jitsu training session, while no changes was observed in LDH activity. Given that these enzymes are located in the cytoplasm of the muscle cell and are not capable of crossing the sarcoplasmatic barrier, increased serum levels of these suggest shedding into the extracellular environment as a result of muscle damage. In an interesting study using two experimental models, fragmented fights and simulated competition, the answers to these enzymes in Jiu-Jitsu fighting were evaluated (Andreato et al., 2013). In the first model the sample was composed by male adult Brazilian Jiu-Jitsu (BJJ) fighters (n = 10) -6 black belts and 4 brown belts- with 10 ± 2 years of experience in the sport. In this model, fights lasting 8min resulted in significant increases of LDH (p = 0,015), while no changes were observed in CK values (p = 0,40) was observed. In the second model, with volunteers with similar characteristics, it was possible to observe the responses of these indirect muscle damage markers in a simulated competition consisting of four matches of 10 minutes, with 20 minutes intervals between fights. At the end of the fourth fight, was observed higher CK values versus pre-fight (p = 0.039), while LDH was not changed during the simulated competition (p = 0.107). In an observational study with BJJ athletes (n = 4) that clashed in the form of competition, with formal structures and

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Financial Support: None.

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Address for correspondence: Hugo Tourinho Filho Avenida Bandeirantes, 3900, Monte Alegre, Ribeirão Preto - SP, Brasil, CEP: 14040-900 E-mail: [email protected] Phone: + 55 16 3315 0348 / Fax: + 55 16 3315 0551

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Received: 14.08.2016 Accepted: 23.10.2016

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