Hormonal response, and women’s competition

ABSTRAC

Hormonal responses to competitions and their relationships have been extensively examined in athletes. The aim of this study was to examine the effect of competition stress on salivary cortisol profiles and dehydroepiandrosterone (DHEA) concentrations in amateur female handball players.
Ten healthy university females’ students, members of a handball club, were selected as  subjects. Salivary samples were collected 30 and 5 min before beginning, between two half times and immediately after four competitions. Salivary DHEA and cortisol were determined by ELISA method.
Salivary DHEA significantly increased during competition(P<0.05), but cortisol and DHEA/cortisol ratio didn’t  significantly changed during and between four competitions (P>0.05).
Since the salivary cortisol level didn’t change during competition in the girl handball players, therefore, in order to know the clear neuro-hormonal responses to competition, using of more suitable stress index is suggested.

HORMONAL RESPONSE,AND WOMEN’S COMPETITION

Introduction

Stress is a biological reaction in response to the external and internal inducers in the body (Selye ea al.,1936 ;Morlyea al.,1991; Chrousos et al.,1992). Recently it has been shown that long term stress has negative impacts on some physiological systems of  the body such as, reproduction (Wingfield2003), growth (Powel et al .,1967), metabolism (Stratakis et al .,1995), immunity (Morlyea al.,1991;  Filaire2001)    and behavior (Chrousos et al.,1992). Since sport competition causes physiological and mental stresses, therefore participating in sport event results in homeostasis disorder (Krueger et al .,2005). On this basis, hormones as the main homeostasis balancing factors are influenced in competitions period. The previous studies have shown that competition in sport setting and non sport setting such as gambling are stressors (Meyer et al .,2000; McCaul et al .,1992). Of course these studies did not investigate the interaction between the winners and losers and the manner of effect of stress on the physiological indices.

The results of these  studies in  two recent decades showed more increase of salivary testosterone in the coin tossing (Hasegawa et al .,2008) and chess winners than the losers(Aizawa et al .,2006), considering the fact that the winners had physical in-activities. Studies on football (Oliveiraa et al .,2009; Carre et al .,2006) hokey (Wang et al .,2009) and basketball (Elias 198116) competitions indicated the same results. The other studies indicating no hormone level difference between the winners and the losers (Filaire et al .,1999). Studies of  Filaire et al. in 1998 and 1999 are the first studies of the kind on the women handball and volleyball players (Filaire et al .,1998; Filaire et al .,2000). These investigations reported a high level of salivary steroid hormones after one competition period. Dehydroepiandrosterone (DHEA) an adrenal specific hormone has anabolic effects on some tissues, by converting to the sexual steroids, like testosterone and estrogen (Aldred et al ., 2009;Mujika et al .,1996).

In women, 90% of circulatory testosterone is derived from the metabolism of environmental precursors particularly of DHEA. Filare et al. (2000) reported that the salivary DHEA acts as a better index than the salivary testosterone in showing the androgens response to exercise in women (Filaire et al .,2000). Cortisol has long term action and is elaborated in stress situation and its secretion is under the influence of adrenocorticotropine hormone (ACTH) secretion in pituitary gland. Any somatic stress ( during physical exercises) or neural (stress due to excitement) induces hypothalamus and secretion of corticotrophin (ACTH) releasing factor, which results in increase of cortisol secretion from the adrenal gland cortex. The secreted cortisol enters the brain cells and its increase or decrease may change the behavioral performance (Krueger et al .,2005; Vervoorn et al .,1991). DHEA/ cortisol ratio is used as an index between the anabolic-catabolic balance and also in evaluating the athlete response to exercise. This ratio is affected by the severity and duration of exercise and any change of this ratio probably are associated to change in athlete’s performance (Kim et al .,2009 ;Chiodo et al .,2009). Catabolic (cortisol) anabolic (testosterone and DHEA) hormones balance has very important application in the performance and recovery periods. The balance between these hormones during heavy exercise and stress condition diminishes and if  recovery to the initial condition is not obtained completely, body secretes cortisol increasingly and reduces DHEA level (Krueger et al .,2005 ;Chiodo et al .,2009). Stress of winning, increases the physical attach and mental stress among players during the match. Also spectators’ crowd, level of expectation from the players and weak result cause mental disturbance in the players, and many of the players  experience it  clearly on the day of competition.

Different factors such as being in risk condition, the  importance of a match, feeling of losing and winning in the players, thinking  of the difficultly of  a match, believing in the role of chance and referee, sexuality, all influence the hormonal response in the winners and losers of  a match (Bateup ea al.,2002).
Therefore, considering the role of stress in the sport competitions and understanding the physiological and mental conditions of the athletes   at the time of match and presence of contraindication about the data on the response, studies on the women hormonal response following competition and need of getting more exact information in designing strategic programs in preventing the increase of stress prior to competition, the present study was designed to investigate the effect of competitive stress on the salivary cortisol and DHEA response in the girl handball players.

Method

Subjects

In this semi -experimental study, 10 girl hand ball players from Sari Branch Azad University participating in the district three Azad University matches held in Chalos Township in 2009, with age 21.5±2.48 years, height 165.67±3.22 cm., weight 58.97±4 kg and body fat percentage 21.35±2.42 were selected as volunteer by simple sampling method.
The criteria for selection were of being healthy, having no history of any mental problem or hormonal disorder, being in similar natural menstrual situation, not using medical drugs and no menstrual and premenstrual pain. All of the participants trained three to four times per week (in all, 8-10 h/week plus 1:30 hour match).

After selection of the subjects for studying, by using questionnaire prepared by the researcher, heart, lung and gland examinations performed by a specialist physician, the objectives and the stages of the research explained to them in detail. Then from each subject written consent was obtained.

Official hand ball competition

All of the subjects participated (two first primary competitions, the third semifinal and the final  competition) in a course of hand ball competition and their salivary samples were collected in each four competitions. These matches were conducted under supervision of district of three Azad University Physical Education Offices in complete official situation observing the world federation rules and regulations.

Prior to each competition the players warmed up for 20 minutes. Since cortisol secretion follows a day night rhythm, the sampling time was the same for all competitions. All matches conducted at 16:00 pm, and continued for 75 minutes. Since the sampling time was the same during the match period, the effect of day night rhythm on hormones secretion was controlled.

Saliva collection

First, the subjects under study washed their mouth, then 3 ml unstimulated saliva was collected into a sterile tube. To determine the basal level variables, the saliva was collected 30 minutes prior to the beginning of the match, also five minutes before, between two half times and immediately after completion of the competition, sent to a clinical laboratory, and  kept at -20 ºC freezer till the examination time. The sampling condition (time and place) was the same for all subjects.

Hormone assay

The salivary DHEA and cortisol were assayed in duplicate, using DEMEDITEC kits, from Germany, referring to the instruction of the company and by ELISA start fax2100 model machine from Awareness Company. The expected normal value of DHEA in women ages 21-30 years is 83-469 pg/ml, the dynamic range is 0-1440 and mean level 206 pg/ml. The sensitivity of kit was 2.18 pg/ml, its functional sensitivity 5.6 pg/ml, and the coefficient of variation (CV) 12.5% (27 Kivlighan2005).
The normal range of cortisol is 50-230 ng/ml (138-635) at 8-10 am and 30-150 ng/ml (82.8-414) at 4 pm, but its dynamic range is 0-800 ng/ml, sensitivity 6.9 ng/ml and the CV 6.5%.

On the day of examination, first the samples were kept in a room with thawing temperature , then, centrifuged at 3000 rpm for 10 minutes for sedimentation of the mucus in the sample. The cortisol and DHEA levels measured using the clear fluid of the supernatant layer. All of the samples were examined in the same condition (time, place and even examiner) condition.

Statistical analyses

The descriptive statistics were  used for analyzing  of means, variances and means changes percentage. Kolmogoroff-Smirnoff test was used for the study of the normality of the obtained data. Analysis of variance (ANOVA) with repeated measurement was used for the evaluation of the hormones level changes in each match. In case of observing any significant difference in determining the place of difference, ordered paired tests (t-dependent) considering the Bonferroni’s P was used. One way analysis of variance and Tukey post hoc test was used for determining the effect of number of matches (four matches) on hormones concentration at the end of one competition period. Value of P<0.05 was considered significant. All of the statistical analysis were done using SPSS-14 software.

Results

Level of salivary DHEA, cortisol and DHEA/cortisol ratio are represented in the fig.1, 2 and 3. The first finding of this study showed that participating in a hand ball competition period has significant effect on DHEA concentration (P=0.001). DHEA concentration in the 1st, 2nd and 3rd matches increased 22%, 3% and 45% respectively indicating statistically insignificant. Increase in the 4th match was 61%, in a way that, DHEA level increased from the beginning and continued immediately after the end of the match. This increase was to the extent that, significant difference (increase) between that of two half times and 30 minutes before (P=0.003 and t=-4.90) and 30 minutes before with after competition (P=0.01, t=-3.67)  was observed. Also DHEA level at the end of four competitions did not show any significant difference (increase) in any of the stages (F=0.493 and P=0.484) Fig.1 section A.
The next finding of the study showed that participating in a period of hand ball competition did not lead to any significant change in the salivary cortisol level (P=0.411). Cortisol level in the 1st, 2nd and 3rd matches increased 58%, 34% and 24% respectively, and decreased in the 4th match 64%, statistically insignificant.

Only increase in the first match was significant, that is to say, cortisol level had significant increase between two periods compared to the resting level (t=-4.21 and P=0.006), but this process reversed till after the competition, which was insignificant. Also cortisol level at the end of four competitions did not show any significant difference (increase) at the end of four competitions (F=1.41, P=0.263), Fig.1 section B.
The final finding of this study showed that participating in a period of hand ball competition did not result in significant change in the DHEA/cortisol ratio (P=0.094). Salivary DHEA/cortisol in the 1st and 2nd matches increased 75% and 32% respectively and decreased in the 3rd and 4th matches 24% and 65% respectively, which was not statistically significant. Also salivary DHEA/cortisol ratio at the end of four competitions did not show significant difference in all of the stages, F=2.29 and P=0.105, Fig.1 section C.

Fig.A- Mean cortisol concentration; B-DHEA;C-Salivary DHEA/cortisol ratio of the subjectsunder study at different stage of sampling* = significant difference to the 30 minutes before competition& = significant difference to 30 minutes before competition,the value based on the mean of standard

Discussion

The obtained data of the present study indicate the nature of multiple hormonal responses in the competition. The reason is related to the vastness of the types of the competitions from the view point of intensity and duration (Filaire et al .,1998; Mujika et al .,1996).
This study showed that participating in a hand ball competition results in increase of salivary DHEA concentration, but no effect on the salivary cortisol level and DHEA/cortisol ratio. Review of the relevant literatures relates the difference of DHEA and cortisol response in competition and training to the different mechanisms of their secretion (Filaire et al .,1998; Filaire et al ., 2000; Kivlighan et al .,2005; Edwards et al .,2006). In case of adrenocorticotrophic hormone (ACTH) deficiency which is a main cortisol secreting hormone, low cortisol is secreted (Edwards et al .,2006). While the adrenal androgens secretion is under minor control of ACTH. This difference between glucocorticoids and adrenal androgens response in the competition could be due to presence of non ACTH and DHEA regulators (Angiotensin , gonadotropin and prolactin) and the other adrenal androgens (Filaire et al ., 2000; Haneishi et al .,2007). Since  salivary samples were used in the present study, no information on the pituitary regulatory system was obtained, while only steroids hormones could be assayed in this method.

Bateup Increase of steroid hormones during match period reported by many reserchers (Bateup et al .,2002;Keizer et al .,1987; Moreira et al .,2009; Kim et al .,2010; Kanalley et al .,1992). Plasma DHEA elevation may be due to combination of factors such as decrease of metabolic clearance (Kim et al .,2010), increase in secretion (Edwards et al .,2006), hematocrit increase, glucocorticoides stimulation (Kivlighan et al .,2005) and hemodynamic difference in the training environment (Kim et al .,2009). Blood volume changes during swimming due to water stress which lead to the changes in hormonal responses (Kim et al .,2009).
Also age of the subjects under study, the method and type of exercise, outcome and duration of the competition account for the similar results. Some workers reported that biochemical pathways, systemic regulation and adrenal androgens secretion differ between men and women. In women the plasma androgens originate from the adrenal cortex (Filaire et al .,2000). Wang et al. (2009) found decrease of DHEA concentration after golf competition in trained females. Some factors such as stage in the menstrual cycle, oral contraceptive or state of hydration may account for this difference (Wang et al .,2009).

Filaire et al. (2000) in the women basket ball and hand ball players and Moreira et al. (2009) on the women football players did not find any significant difference on the salivary DHEA and cortisol concentration following participating in the competition (Filaire et al. ,2000; Moreira et al. ,2009).

Kim et al. (2010) reported significant increase of cortisol concentration after a rugby match. In a way that, the subjects competed in field and in laboratory (with similar condition). The cortisol level did not change significantly in laboratory condition. Psychological factors influence cortisol response to competition (31 Kim et al. (2010). The probable disagreement may be that the previous studies were done in elite male and female in laboratory condition or on the runners and cyclists.
Also time of sampling played part in obtaining different results. In the present study sampling was done in four stages (30 min before, immediately before, between two half times and at the end of competition), but in the previous studies the salivary sampling was done only before and immediately after one winning match and one losing match (Kim et al. ,2010), before and immediately after one competition period of 16 weeks (Krueger et al.,2005).

In the present study considering the type of activity model which comprised continued running and quick movement (Elias .,1981), shows  that from the view point of bodily condition, the subjects did not tolerate high body stress and the intensity was not that much to stimulate cortisol secretion. In men, free testosterone/cortisol ratio (FT/C) and testosterone /cortisol (T/C) ratio are used as an index of adaptation of training (Kim et al .,2009). In females parameters of this hormone are used with more precautions due to particular profile of this hormone, which extensively has relation with the menstruation cycle and use of oral contraceptive  (Kim .,2009; Chiodo et al 2009). Some reports indicate that cortisol or adrenal androgens (DHEA and Δ4 androstandions) are not under influence of different menstrual stages (Vervoorn et al .,1991). While other reports show  the effect of menstrual stage on this hormone (Kanalley et al .,1992 ;Genazzani et al .,1983).

In the present study none of the study subjects used oral contraceptive and all of them had regular menstrual cycle.
Considering the elevation of DHEA and lack of change in cortisol concentration, no change has been observed in DHEA/cortisol ratio. Majika et al. (1996) found that DHEA/cortisol ratio has negative relation with number of the  swimming training weeks (Majika et al. 1996). This finding  was approved  by further research with the data given by Filaire et al (1999); Bouget et al (2006); Doan  et al. (2007); Edwards et al. (2006), disagree with the data obtained in our study, which could be due to the type of sport, training environment, sex age of the subjects under study and also difference in the test methodology (Filaire et al (1999); Bouget et al (2006); Doan  et al. (2007); Edwards et al. (2006)). In all, the results of the present study showed  that a period of handball competition leads to elevation of androgens concentration and no change in glucocorticoids concentration. Therefore adrenocortical metabolism is not under influence of competitive stress. Since neural system response was not measured in this study (salivary measurement) which is the limitation of this study, knowing the pituitary regulatory system was impossible. Therefore it is proposed that in the future studies the relevant factors with the adrenocortical response (neural factors)should be taken into consideration.

ACKNOWLEDGMENTS

Thanks to the research deputy of Islamic Azad University Sari branch for providing the finance, and to the laboratory staff for their cooperation in performing the tests.

REFERENCES

Selye ,H.A. (1936): Syndrome produced by diverse nocuous agent. J Neuropsychiatry Clin Neurosci,10(2):230-231.

Morly ,J.E., Benton, D., Solomon ,G.F.(1991): The role of stress and opioids as regulators of the immune response. In: Stress, Neuropeptides, and Systemic Disease. MCCubbin :Academic,5:221-231.

Chrousos, G.P., Gold ,P.W. (1992): The concepts of stress and stress system disorders Overview of physical and behavioral homeostasis. JAMA,267(9):1244-1252.

Wingfield ,J.C., Sapolsky ,R.M.(2003: ( Reproduction and resistance to stress: when and how. J Neuroendocrinol,15(8):711-724.

Powel, G.F., Brasel, L.A ., Raiti ,S., Blizzard ,R.M .(1967): Emotional deprivation and growth retardation simulating idiopatic hypopituitarism. II. Endocrinologic evaluation of the syndrome. N Engl J Med,267(23):1279-1283.

Stratakis ,C.A., Chrousos ,G.P. (1995): Neuroendocrinology and pathophysiology of the stress system. Ann NY Acad Sci,771:1-18.

Filaire ,E., Sagnol ,M., Ferrand ,C. (2001):Psychophysiological stress in judo athletes during competitions. J Sports Med Phys Fitness,41(2):263-268.

Krueger, T.H., Schedlowski, M., Meyer ,G. (2005): Cortisol and heart rate measures during casino gambling in relation to impulsivity. Neuropsychobiology,52(4):206-211.

Meyer, G., Hauffa ,B.P., Schedlowski, M., Pawlak ,C., Stadler, M.A ., Exton ,M.S. (2000):Casino gambling increases heart rate and salivary cortisol in regular gamblers. Biol Psychiatry48:948-953.

McCaul, K.D., Gladue, B.A., Joppa ,M. (1992): Winning, losing, mood, and testosterone.
Horm    Behav  ,26(4):486-     504 .

Hasegawa ,M., Toda ,M., Morimoto, K. (2008):Changes in salivary physiological stress markers associated with winning and losing. Biomed Res,29(1):43-46.

Aizawa, K., Nakahori, C., Akimoto ,T. (2006): Changes of pituitary, adrenal and gonadal hormones during competition among female soccer players. J Sports Med Phys Fitness,46(2):322-27.

Oliveiraa ,T., Gouveiaa, M.J., Oliveiraa, R.F. (2009): Testosterone responsiveness to winning and losing experiences in female soccer players. Psychoneuroendocrinology,34(7):1056-1064.

Carre ,J., Muir, C., Belanger, J .,Putnam ,S.K. (2006): Pre-competition hormonal and psychological levels of elite hockey players :relationship to the “home advantage” .Physiol Behav,89(3):392-398.

Wang ,H.T., Chen, S.M., Lee ,S.D., HSU, M.C. , Chen, K.N ., Liou, Y.F. , Kuo ,C.H.(2009): The role of DHEA-S in the mood adjustment  against negative competition outcome in golfers.J Sports Sci,27(3):291-297.

Elias ,M. (1981): Serum cortisol , testosterone and testosterone –binding globulin responses to competitive fighting in human males. Aggressive  Behav,7: 215-224.

Filaire, E., Scanffc, L.E., Duche, P., Lac, G . (1999):The relationship between salivary adrenocortical hormones changes and personality in elite female athletes during handball and volleyball competition .Res Q Exerc sport,70(3):297-302.

Filaire ,E., Duche, P., Lac, G. (1998):Effects of amount of training on the saliva concentration of cortisol ,  Dehydroepiandrosterone and on the Dehydroepiandrosterone: cortisol concentration ratio  in woman over weeks of training. Eur J Appl Physiol Occup Physiol,78(5):466-471.

Filaire, E., Lac, G. (2000):Dehydroepiandrosterone (DHEA)  rather than testosterone shows saliva androgen responses to exercise in elite female Handball players. Int J Sports Med,21(1):17–20.

Aldred, S., Rohalu ,M., Edwards, K ., Burns, V. (2009): Altered DHEA and DHEAS response to exercise in healthy older adults. J Aging Phys Act,17(1):77-88.

Mujika, I., Chatard, J.C., Padilla, S.(1996): Hormonal responses to training and its tapering off in competitive swimmers. Eur J Appl Physiol Occup Physiol,74(4):361-366.

Vervoorn ,C., Quist, A.M., Vermulst, L.J.(1991): The behavior of the plasma free testosterone/cortisol ratio during a season of elite rowing training. Int J Sports Med,12(3):257-263.

Kim ,K.J., Chung, J.W., Park ,S .,Shin, J.T.(2009(:Psychophysiological Stress Response during Competition between Elite and Non-elite Korean Junior Golfers. Int J Sports Med,30(7):503-508.

Chiodo ,S., Tessitore, A., Cortis ,C. (2009): Stress-related hormonal and psychological changes to official youth Taekwondo competitions. Scand J Med Sci Sport, 44(1): 61-66.

Bateup, H.S., Booth ,A., Shirtcliff ,E.A., Granger, D.A. (2002):Testosterone,cortisol and women’s competition. Evol Hum Behav,23:181-192.

Kivlighan, K.T., Granger, D.A., Booth, A. (2005): Gender differences in testosterone and cortisol response to competition. Psychoneuroendocrinology,30(1):58-71.

Edwards, D.A., Wetzel, K., Wyner ,D.R. (2006): Intercollegiate soccer: Saliva cortisol and testosterone are elevated during competition, and testosterone is related to status and social connectedness with teammates. Physiol Behav, 87(1):135-143.

Haneishi ,K., Fry, A.C., Moore, C.A.(2007): Cortisol and stress responses during a game and practice in female collegiate soccer players. J Strength Cond Res,21(2):583-588.

Keizer, H.A., Kuipers ,H., de Haan ,J . (1987): Effect on a 3-month endurance training program on metabolic and multiple hormonal responses to exercise. Int J Sports Med,8 Suppl 3:154-160.

Moreira, A., Arsati, F., de Oliveira Lima Arsati ,Y.B.(2009): Salivary cortisol in top-level professional soccer players. Eur J Appl Physiol,106(1):25-30 .

Kim, K.J., Park, S., Kim ,K.H.(2010):Salivary cortisol and immunoglobulin A responses during golf competition vs. practice in elite male and female junior golfers. J Strength Cond Res,24(3):852-58.

Kanalley, J.A., Bioleau ,R.A., Bahr, J.M. (1992): Cortisol levels during prolonged exercise: the influence of menstrual phase and menstural statua. Int J Sports Med,13(4):332-336.

Genazzani, A.R., Inaudi ,P.D., Ambrogio, G.(1983): Plasma androgens and menstrual cycle: physiopathological correlates from adolescence to menause. Horm Res,18(1-3):84-97.

Bouget ,M., Rouveix, M., Michaux ,O.(2006):Relationship among training stress, mood and DHEA sulphate cortisol ratio in female cyclists .J Sports Sci,24(12):1297-302.

 Doan , B.K.and Newton ,R.U., Kraemer, W.J.(2007):Salivary cortisol, testosterone, and T/C ratio responses during a  36–hole golf competition. Int J Sports Med,28(6):470-479.