International Journal of Basic Sciences & Applied Research. Vol., 4(1), 38-43, 2015 Available online at http://www.isicenter.org ISSN 2147-3749 ©2015 The Effect of Endurance Training on the Ratio of Serum Cortisol to Dehydroepiandrosterone (DHEA) in Inactive Young Women Somayeh Mahdaviyan1*, Laleh Behboodi1, Mojtaba Ezadi2 1 Department of Physical Education, Islamic Azad University, Islamshahr Branch, Tehran, Iran Department of Sports Sciences and Physical Education, Saveh Branch, Islamic Azad University, Saveh, Iran 2 * Corresponding Author Email: [email protected] Abstract The aim of present study was to determine the effect of endurance training on balance of anabolic to catabolic in inactive young woman's body (Ponjee, 1994). Inactive young woman aged 24.50 ± 2.653 years, height 164.64 ± 5.624 cm, weight 57.928 ± 4.921 kg, body mass index (BMI) 21.376±1.031 kg by the square of height the meter, selected through purposive convenience sampling and randomly divided into exercise and control groups (each group, n=10). Practice group participated at 8 weeks of endurance training for 3 days a week with highly increasing and the control group did not follow any regular physical activity, and only performed the normal and routine activities. Before and after training, was taken blood samples from subjects of and measured and calculated cortisol, DHEA and ratio of serum cortisol to DHEA. T-test used for comparison of variables. Cortisol in the exercise group compared with the control group was significantly increased (P<0.05). Also the rate of DHEA and ratio of serum cortisol to DHEA in the exercise group compared with the control group significantly decreased. 8-week endurance training similar to the present protocol providing a catabolic environment in the body of the inactive young women. However, it appears that the results of this study is due to the overtraining, and besides the psychological and physiological condition of regnant the subjects may be according to the protocol of exercises, apply high training intensity in the final weeks of exercise, especially for those who were disabled before it can be considered overtraining. Keywords: Endurance training, Ratio of DHEA to cortisol, DHEA, Serum cortisol. Introduction One of the most important methods for evaluating the intensity of the exercise and its effect on athletes is measure biochemical variables, In this regard, biochemical variables can be divided into three general categories of enzymes, hormones and metabolites. Among the hormones more concerned to anabolic and catabolic hormones, if so that catabolic hormones increase subsequent exercise was signs of high pressure of activity or if the balance be established between anabolic and catabolic hormones athlete is in good fitness condition. In this regard, testosterone and cortisol hormones to order as anabolic and catabolic hormones are considered more than others hormones and the ratio of these hormones is another useful indicator for determining the condition and pressure of work and training (Obminski, 1997). Endurance training attention too many of people, especially women. This exercise causes physiological changes and adaptations that most of it can be found in the neuromuscular system and hormones. Measuring the indexes of pressure of practice followed by various training programs can contribute to a better understanding of the acute and chronic effects of endurance training. For this purpose, in men used testosterone to cortisol ratio and said that if this ratio decrease 30% or greater, indicate that overtraining syndrome, But in women because of this hormone secreted by the ovaries and adrenal gland being used from another anabolic hormone that is the precursor of testosterone and called DHEA. So in women DHEA to cortisol ratio as an indicator of exercise pressure (Urhausen, 1995). In this circumstance the level of cortisol is dramatically increased that due to exercise stress, lack of adequate rest and nutrition inappropriate, and it is well known energy reserves are not recovering well and retrieved, and in the next training athlete 38 Intl. J. Basic. Sci. Appl. Res. Vol., 4(1), 38-43, 2015 earlier exhausted and function is impaired (Obminski, 1997). Now, if athlete after exercise, has enough rest or good nutrition lower this ratio decreases. On the other hand athlete who suddenly increase his exercise volume and intensity, exposure decrease of ratio. Recently endurance training take into consideration by many of women that has effects for health and fitness. If the exercise pressure was excess capacity of physiological not only improve performance but impair the performance, and puts a person under the effects of overtraining. On the other hand, if the exercise pressure was low, is not created significant improvements in performance and physiological adaptations. Methodology Subjects The population of this research were consisted of all inactive healthy girl student of Azad University of Tehran center (20 to 28 years old). Recall over 20 person have declared their readiness to participate in research. The number of 20 inactive student girls (20 to 28 years old) 24.50 ± 2.653 years, height 164.64 ± 5.624 cm, weight: 57.928 ± 4.921 kg and body mass index (BMI) 21.376) ± 1.031 kg per cubic meter, selected through purposive convenience sampling and randomly divided into exercise and control groups (each group, n = 10). All subjects had a complete physical health (physician confirmation). Researcher with subjects Homogenization (exception genetic material), reduce probability of influence disruptive variables on dependent variables. Demographic characteristics of the subjects are presented in Table 1. Subject of two groups were similar in terms of age, height, weight and BMI (P> 0.05). Table 1. Demographic characteristics of subjects. Group Exercise Control Total Age 2.636±24.571 24.428±2.878 2.653±24.50 Height 4.790±165.43 163.86±6.644 5.624±164.64 Weight 58.571±5.503 57.285±4.608 4.921±57.928 BMI 21.412±1.488 21.34±0.295 21.376±1.031 Methods of data collection One week before run study, subjects were familiar with exercise protocol in justification meeting. Also the personal and demographic characteristics of the subjects were measured. Then, were taken blood samples, 48 hours before the start of the test in the fasting state, the subjects conducted their exercise program for 8 weeks increasingly. At this time the control group did not perform any physical activity and sport, and only do their daily activities. After completing the 8 weeks of training, and after the proportional rest with gap of first day of sampling and the start of training (48 hours), second test session done like to first test and obtained rest blood samples in the fasting status. Exercise program Endurance training was consisted of 8 weeks and 3 days each week. A percentage of maximum heart rate and duration of exercise was considered as intensity and volume of training. Each run session was 35 minutes including a 5 minute warm up, 25 minutes main exercise and 5 minutes cool-down? 25 minutes of main exercise was include running on the treadmill. For first to eight weeks done with 60%, 65%, 70%, 70%, 75%, 75%, 80% and 85% of maximum of heart rate. To display the heart rate on the treadmill screen was used the belt. Each subject started and finished all activities meetings at own time, that this time were the same for all. Hormonal analysis: 48 hours before and 48 hours after training, were collected blood sample of subjects of both groups from central venous at a rate of 5 ml. It should be noted that after each session, participants were considered to drink enough fluids to compensate for lost fluid. Blood samples was poured in sterile tubes containing K3EDTA, heparin and EDTA tubes were placed in ice and then for a few minutes left at room temperature, followed by centrifugation was separated serum from plasma for 10 minutes at around 3500 RPM. All blood samples were kept to be frozen at -20 ° C until to reach the laboratory, and there began laboratory test immediately. Serum cortisol level was measured for each sample from using the ELISA way and IBL kit with sensitivity of 2.5 ng ml, also DHEA was measured for each sample from using the ELISA way and IBL kit with sensitivity of 0.108 ng ml. To calculate the ratio of cortisol to dehydroepiandrosterone, after the conversion of unit of both hormone to nanomole per liter and also was used to formula of ng/ml * 275.9 = nmol/l for conversion unit of cortisol and formula of ng/ml * 3.47 = nmol/l for conversion unit of dehydroepiandrosterone. Statistical Methods: First the variables were described with the mean and standard deviation. Then was used Smirnov Kolmogorov test for the determine of normal distribution and license to use the parametric or non-parametric tests. Also was used paired t-test to investigate the changes of variables in the exercise and control groups, also was used independent t-test in order compared to the values of each variables between the exercise and control groups. For all statistical tests was set the significance level equal to 0.05, also was used the statistical software SPSS version 16 for statistical calculations. 39 Intl. J. Basic. Sci. Appl. Res. Vol., 4(1), 38-43, 2015 Results Statistical description of the variables is presented in table 2. Results of dependent and independent t-test (respectively to assess intragroup and intergroup comparisons of variables) are presented in tables 2 and 3. Weight and BMI decreased in the exercise group compared with the control group (P <0.05). Cortisol increased in the exercise group compared with the control group (P <0.05). DHEA and DHEA to cortisol ratio also decreased in the exercise group compared with the control group (P> 0.05). Table 2. Mean and standard deviation of variables. Variables Groups Training Control Training Control Training Control Training Control Training Control Weight (kg) BMI (kg/m2) Cortisol (ng/ml) DHEA (ng/ml) DHEA to Cortisol Ratio Pre 58.571±5.503 57.285±4.608 21.412±1.488 21.34±0.295 29.072±26.153 19.407±8.884 92.228±18.286 100.23±19.144 5.950±6.005 6.434±3.414 Post 54.428±5.912 57±4.932 19.870±1.363 21.230±0.557 31.837±26.837 19.45±8.988 85.157±16.411 100.57±20.562 4.474±3.928 6.524±3.702 Table 3. Results of t-tests to examine within-group variables. Variables Weight (kg) BMI(kg/m2) Cortisol (ng/ml) DHEA (ng/ml) DHEA to Cortisol Ratio Groups Training Control Training Control Training Control Training Control Training Control t 6.96 0.67 6.28 0.67 10.71 1.96 5.19 1.29 11.06 2.28 df 9 9 9 9 9 9 9 9 9 9 Sig. 0.000 * 0.52 0.001 * 0.52 0.000 * 0.08 0.001 * 0.22 0.000 * 0.04 * * Significant in P≤0.05 Table 4. Results of t-test for comparison between groups of variables. Variables Weight (kg) BMI(kg/m2) Cortisol (ng/ml) DHEA (ng/ml) DHEA to Cortisol Ratio * Significant in P≤0.05 ∆: Changes before and after exercise 40 Groups Before After ∆ Before After ∆ Before After ∆ Before After ∆ Before After ∆ t 0.47 0.88 5.29 0.12 2.44 5.04 1.47 2.27 6.93 0.52 2.43 3.01 3.46 2.07 5.89 df 12 12 12 12 12 12 12 12 12 12 12 12 12 12 12 Sig. 0.64 0.39 0.000 * 0.90 0.03 * 0.000 * 0.15 0.03 * 0.000 * 0.61 0.02 * 0.007 * 0.003 * 0.053 0.000 * Intl. J. Basic. Sci. Appl. Res. Vol., 4(1), 38-43, 2015 Discussion and Conclusion Based on research findings, endurance training caused to increase serum cortisol levels significantly. In line with the findings of this present, Hossieni et al (2009), also reported cortisol increases in the endurance training group, but the Aghaalinejad et al (2013) showed that when resistance training implemented as endurance and consistently training, can reduce serum cortisol in young women during 8 weeks. Also in Hiruntrakul and Ratanavadee (2010), showed cortisol levels of rest had not significant difference between endurance training and control groups, they concluded that 12 weeks of moderate-intensity and low repetition don have effect on cortisol in inactive people. In Hejazi and Attarzadeh Hosseini (2010) Thirteen elite male runners participated in a semi-endurance exercise program consisted of 14 weeks and 12 sessions per week (morning and afternoon), blood samples were collected in three stages, before preparation, after preparations and before the competition. Cortisol decreased significantly in the after stage of preparation, although increased in stage of before competition. Unlike the present findings, Chatard (2002) reported that non-athletes are more cortisol concentration than athletes in rest. Different results may be due to differences in the exercise protocols. The present results suggest that exercise increases resting cortisol levels. The researchers stated that Cortisol has stressful role as a hormone, due to hypothalamic-pituitary-adrenal axis stop will be used as a valid indicator for the diagnosis of overtraining and prevention of mental increasing activities acquired of the tournaments. Two factors, intensity and duration of exercise change the response of the adrenal pituitary-hypothalamic axis to exercise. It’s possible in the study that training intensity up to 80 or 85% has partly caused overtraining in maximum rate at end session. Recent studies have shown that it is better to practice with a uniform intensity in the final weeks of rehearsal as the last 3 or 4 weeks and impose additional burden not done with higher intensity in sessions before the sampling, so this leads to some kind of consistency. Other factors that alter the cortisol response to exercise include Hyper Hydration, food and the circadian rhythm. Physiological loads (intensity and volume) non-proportional and intensities exercise leads to increase cortisol due to raising the secretion of the adrenocorticotropic hormone that is a response to increased susceptibility to stress on the hypothalamic-pituitary-adrenal axis. Different workloads (intensity and volume) different exercises (strength, endurance, power) micro cycle training courses, macro cycle and individual compatibility show different hormonal responses. Since the present study showed an increase in cortisol, it can probably be due to the incompatibility of the hypothalamic - pituitary-adrenal axis to training conditions and lack of lowering cortisol sensitive and lack of sensitivity change of tissue to the cortisol. On the other hand, the results suggest that physical activity cause increasing the levels of releasing hormone of catecholamines and cortisol . The time of increasing or their achieving to a base level is not same on the all of them. Some of them, such as cortisol unlike the catecholamines have slower fluctuations will affect the physiology of the body for a few days. Based on the results of this study, slow drop of cortisol which lasts for several days is one possible reason for the increase in cortisol levels. This means that if the second sampling was performed 72 hours after the last training session insteade of 48 hours after the last session, this result was not gained because disabled people spend a longer recovery period than active people. Exercise load presented in this study for an ordinary athlete and for a normal and non-active person may be too according to the principle of individual differences. Therefore, based on the findings of overtraining can effect on anabolic and catabolic steroids such as cortisol (Khaledan, 2002). Some studies have considered the performance. However, there are conflicting results in this area and as mentioned above, it can be related to the nature of the sport, the exercise volume and intensity, measurement interval, subjects, the sampling manner( blood, saliva )and the willingness of individuals(Engelmann, 2004). Numerous studies have been conducted on factors of hormones overtraining, and overtrainings has caused an increasing IL-6 and this leads to increase lipolysis and thus it stimulates cortisol and other cytokines (Asgarpour & Nazarali, 2012). In the present study, lose weight and body mass index may also show an increase in IL 6 which is not measured. Researchers have expressed that the lack of hormonal changes may be the result of appropriate and normal exercises with proper recovery thereby cause to maintain the stability the hypothalamic-pituitary axis. Thus the appropriate recovery allows adaptation to stress that occurs during the stress response (Halson & Jeukendrup, 2004). However, the increase in cortisol in this study may come from the improper training or lack of proper recovery for this group. Based on research findings, one endurance training period significantly reduce the levels of DHEA of serum. Hosseini et al (2009) also reported a decrease in DHEA in the endurance training group. In contrast, Sato (2013) reported that DHEA levels, 5 alpha Dihydrotestosterone and5-alpha reductase Dihydroergotamine were significantly higher in skeletal muscle of diabetic rats after 6 weeks in exercise group. Also Johansson and Lars-Eric (2013) reported that the DHEA sulfate and the ratio of DHEA sulfate to cortisol were significantly higher than in amateur hockey players to professional hockey athletes but it was not like this in the case of cortisol. Aghaalinejad (2013) in their study reported an increasing the DHEA serum of young women during 8 weeks. In Hiruntrakul and Ratanavadee (2010), resting levels of total testosterone and free testosterone showed no significant difference between the two endurance training and control groups. They concluded that 12 weeks of exercising with moderate-intensity and low repetition have no effect on testosterone of disabled people. In Hejazi and Attarzadeh 41 Intl. J. Basic. Sci. Appl. Res. Vol., 4(1), 38-43, 2015 Hosseini (2010) testosterone after preparation and before the non-compete period was increased and decreased, respectively. Hakkinen and Pakarinen (2005), showed respectively, after 12 and 21 weeks of strength and endurance training observed the increasing the amount of saliva di DHEA on disabled women. Arce and De Souza (1993) found that subjects of endurance and resistance training have lower levels of testosterone than control group. In contrast to the present findings, Keizer (1989) reported that the DHEA sulfate dioxide increases in response to endurance exercise. Important characteristic of overtraining syndrome has listed reduction of function as a result of an imbalance between anabolic and catabolic hormones with dysfunction in the hypothalamus-pituitary-adrenal axis.The role of testosterone as an anabolic hormone, and cortisol is important as a catabolic hormone. Testosterone on hypertrophy muscle and muscle glycogen synthesis is important and indicates the anabolic state in the body. According to the researchers’ results, the long-term exercises increase plasma cortisol levels and reduce their testosterone (Hakkinen & Pakarinen, 2005; Arce & De Souza, 1993; Keizer, 1989). Perhaps the time of practice of the present study was also long to prepare participants. DHEA of blood can turn to Androstenedione, which can then be converted to testosterone. Researchers have considered reduction of testosterone levels in the blood a key indicator of fatigue and overtraining (Katsuji, 2010; Ponjee, 1994). Resting testosterone affected by several factors such as exercise varies (duration and intensity), individual characteristics (age, sex) and diet. Testosterone reduction after exercise induced increases in intensity, duration, repeat action the exercise load that do not give the opportunities to restoration and reset (Platen, 2002). Testosterone levels can also reduce followed by intense endurance exercise and resistance exercise and particularly when is associated with dietary restriction and negative energy balance (Engelmann et al., 2004). In the present study the weight reduction and BMI of negative energy balance caused by diet may also not be controlled. Mechanisms of reduction of testosterone levels may be because of gonad releasing hormone secreted by the hypothalamus, prolactin increasing and inhibition of by the LH hormones secreted of pituitary or directly inhibition by cortisol (Halson & Jeukendrup, 2004). Also increase in testosterone levels is mainly depends on the done work load, involved muscle mass, and rest between repetitions and athletes preparation (Viru, 2005; Kraemer, 1988) said resting concentration of testosterone levels may be a reflection of the current state of muscle tissue and whether establish the anabolic conditions or not. In this present study also levels of DHEA reduced. This issue could be indicative of catabolic conditions. The results demonstrate that this reduction may relate to non-adjusted for action on the hypothalamic-pituitary axis on the adrenal and ovary in response to the balance between the exercise activity and recovery or intensity and volume of exercise (Pederson, 2000). On the other hand, DHEA has been used empirically in patients with depression and has been successful in improving depression and memory. So regard to non- control the psychological conditions of subjects may have changed estrogen rates due to mental conditions. Based on research findings, one endurance training course significantly reduced levels of (DHEA) compared to serum cortisol. Hosseini et al (2009) also reported the increase in de (DHEA) to cortisol in the endurance training group. Johansson and Lars-Eric (2013) reported that the sulfate of (DHEA) and ratioof DHEA sulfate to cortisol was significantly higher in amateur hockey players to professional hockey players. Aghaalinejad (2013) in their study demonstrated an increase ratio of DHEA to serum cortisol during the 8 weeks. Hejazi and Attarzadeh Hosseini (2010) showed that the testosterone has significantly increase compared to cortisol. Consistent with our findings, Ponjee (1994) also has reported a significant decrease in DHEA and cortisol after long-term endurance activities. DHEA ratio to cortisol is used as a marker of training stress in athletes. This ratio is influenced by the intensity and duration of exercise, any change in this ratio can be associated with a probably change in immune function (Hossieni & Aghaalinejad, 2009). Research shows that DHEA ratio reduced to cortisol, in the individuals who are suffering from overtraining (Halson & Jeukendrup, 2004; Moore & Fry, 2007). In this study, we observed a significant decrease in the ratio of these two hormones in the control and experimental groups. This decrease reflects the imbalance of the anabolic/catabolic and creating conditions of catabolic in the training phase of the research group. Mainly different in the various results may be due to differences in the type of training and the individual's physical and psychological characteristics. The results show a lack of adaptation of overtraining to the stress, training conditions and are a sign of an appropriate return to its original state and fatiguecausing factors (Hayes et al., 2010). It can also be stated since the subjects in this study(inactive women)has low preparation levels, changes in the measured parameters affect by exercise can probably be attributed to their low fitness, perhaps the exercise protocol in individual with better fitness will lead to overtraining. Based on research findings, such it can be concluded that the 8-week endurance training creates a catabolic environment in the non-active young woman's body and this practice can lead to weight loss due to hormonal changes and are useful for people who need to lose weight. But it seems the results of the research indicate overtraining and furthermore, physiological and psychological conditions of the study may be considered a form of overtraining according to the exercise protocol of implying increase the training intensity in end weeks especially for those who have already inactive. 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