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Changes of stress proteins and oxidative stress indices with progressive exercise training in elderly men
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ORIGINAL ARTICLE
Changes of stress proteins and oxidative stress indices with progressive exercise training in elderly men Changements des protéines de stress et des marqueurs du stress oxydant lors d’un entraînement progressif chez des sujets âgés de sexe masculin S. Atashak a,∗, K. Azizbeigi b, M. Ali Azarbayjani c, S.R. Stannard d, F. Dehghan e, R. Soori e a
Department of Exercise Physiology, Mahabad Branch, Islamic Azad University, Mahabad, Iran Department of Exercise Physiology, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran c Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University, Tehran, Iran d School of Sport and Exercise, Massey University, New Zealand e Department of Exercise Physiology, Faculty of Physical Education and Sport Sciences, University of Tehran, Tehran, Iran b
Received 8 December 2015; accepted 16 January 2017
KEYWORDS Malondyaldeyde; Heat shocks protein-70; Carbonyl protein; Total antioxidant capacity; Elderly men
∗
Summary Purpose. — Excessive generation of free radicals and oxidative stress play an important role in acceleration of the aging process and contribute many chronic diseases. Nevertheless, it has been shown that the elderly who are physically active benefit from exercise-induced adaptation in cellular antioxidant defense systems and associated increase in the generation heat shocks proteins (HSPs). Therefore, the aim of the present research was to investigate the effects of 14 weeks participation in concurrent (resistance and aerobic) training on indices of oxidative stress and HSP70 concentration in inactive elderly men.
Corresponding author. E-mail address: [email protected] (S. Atashak).
http://dx.doi.org/10.1016/j.scispo.2017.01.006 0765-1597/© 2017 Elsevier Masson SAS. All rights reserved.
Please cite this article in press as: Atashak S, et al. Changes of stress proteins and oxidative stress indices with progressive exercise training in elderly men. Sci sports (2017), http://dx.doi.org/10.1016/j.scispo.2017.01.006
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S. Atashak et al. Methods. — Twenty-four inactive elderly men voluntarily participated in this research and were assigned to either training (n = 12) or control (n = 12) groups. In the control group, participants were advised to maintain their normal lifestyle during the study, while the training group was prescribed a combination of resistance and aerobic training for 14 weeks. At baseline and after 14 weeks, venous blood samples were obtained for measurement of malondialdehyde (MDA) and carbonyl protein (PC), total antioxidant capacity (TAC) and HSP70 concentration. Results. — The results indicate very significant effects of exercise training on all measured experimental variables when compared to the control group: MDA and PC concentrations decreasing, whilst HSP70 and TAC in aged men were increased significantly after exercise training in training group (all P < 0.01). Conclusion. — This study shows conclusively that concurrent exercise training for 14 weeks decreases indices of oxidative stress in elderly men and can ameliorate age-related deficits in HSP70 and the circulating antioxidant defense system. © 2017 Elsevier Masson SAS. All rights reserved.
MOTS CLÉS Malondyaldéhyde ; Protéine HSP 70 carbonyle ; Capacité antioxydante totale ; Sujet âgé
Résumé But. — Une production excessive de radicaux libres et le stress oxydant jouent un rôle important dans l’accélération du processus de vieillissement et sont impliquées dans beaucoup de pathologies chroniques. Néanmoins, il a été démontré que les personnes âgées qui sont physiquement actives profitent des modifications induites par l’exercice dans les systèmes de défense antioxydants et l’augmentation associée des protéines du choc thermique (HSP). Le but de ce travail était donc d’étudier les effets de la participation à 14 semaines d’entraînement associé (résistance + endurance) sur les marqueurs du stress oxydatif et les concentrations de HSP70 chez des hommes âgés sédentaires. Méthodes. — Vingt-quatre hommes âgés sédentaires ont volontairement participé à cette recherche 20 et ont été affectés aux groupes entraînement (n = 12) ou contrôle (n = 12). Dans le groupe témoin, les participants ont été invités à maintenir leur mode de vie habituel au cours de l’étude, tandis que le groupe entraînement a réalisé 14 semaines d’entraînement associé (résistance + endurance). Avant le début de l’étude et à la fin des 14 semaines, des échantillons de sang veineux ont été obtenus pour la mesure du malondialdéhyde (MDA), de la protéine carbonyle (PC), de la capacité anti-oxydante totale (TAC) et de la concentration en HSP70. Résultats. — Les résultats montrent un effet marqué de l’entraînement physique sur toutes les variables mesurées en comparaison avec le groupe contrôle : les concentrations de MDA et PC diminuent, la ue HSP70 et la TAC augmentent de fac ¸on significative après l’entraînement physique dans le groupe de sujets âgés soumis à l’entraînement (p < 0,01). Conclusion. — Cette étude montre de fac ¸on concluante que 14 semaines d’entraînement combinant résistance et endurance diminuent les marqueurs de stress oxydant chez des sujets âgés de sexe masculin et peuvent ainsi contribuer à corriger les déficits en HSP70 et en système de défense antioxydant liés à l’âge. © 2017 Elsevier Masson SAS. Tous droits r´ eserv´ es.
1. Introduction Aging, as a complex multifactorial biological process, is a progressive decline in physiological function that is often accompanied by increased susceptibility to diseases leading to disability and often premature death [1]. The free radical theory of aging, involving the accumulation of deleterious effects caused by reactive oxygen species (ROS) and reduced antioxidant defenses in elderly people, is the most popular theory proposed for the aging process [2]. According to this theory, oxidative stress increases with age and the ability of organism to cope with cellular damage induced by this stress decrease [3], finally causing cellular dysfunction and resulting in apoptosis [4]. Moreover, abundant experimental and observational studies have found that age-associated oxidative damage and reactive oxygen species (ROS) have
an important role in diverse chronic age-related diseases such as cancers, cardiovascular disease (CVD), Alzheimer’s and Parkinson’s diseases [5,6]. It seems that proteins, because they have specific functions, are one of the most obviously affected macromolecules in age-related oxidative damage [7]. An age-related increase in the protein carbonyl (PC) concentration, which has been used as a marker of ROS-mediated protein oxidation, was observed in various tissues [8]. Mutlu-Türko˘ glu et al., in this regard, observed that plasma malondialdehyde (MDA) and PC levels associated with accumulation of oxidative damage were significantly higher in the plasma of elderly subjects as compared to young subjects [9]. Moreover, it has been shown that the capacity to produce heat shock proteins (HSPs) decreases with aging in normal older subjects [10]. HSPs act as molecular
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Effect of exercise training on oxidative stress chaperones that regulate the conformation and functions of a large number of cellular proteins and has been demonstrated that to protect cells against oxidative stress injury [11]. Indeed HSPS, and especially Hsp70, may be very important in normal physiological and stressful (pro-oxidative) conditions [10]. A sedentary lifestyle typical of the elderly is a key factor in the etiology and progression of age-related chronic disease [12]. However, the majority of published data suggests that regular exercise training improves the health markers and functional capacity in older people and has considerable efficacy counteracting the aging process and associated diseases [13]. It is plausible that elderly people who engaged in physical activity, besides improving of strength, endurance and subsequently functional capacity, benefit from exerciseinduced adaptation in cellular antioxidant defense [14]. In this context, it has been reported that long-term physical training increases the activity of antioxidant enzymes and HSPs expression, and therefore reduced markers of oxidative damage [15]. In this regard, while there are indications that HSP70 expression decreases with age, long-term exercise training reduced apoptosis rate and significantly increase HSP70 in the cardiac muscles of training animals compared to sedentary control animals [16]. Further, it has also been reported that 12 week low-volume of physical activity significantly reduces the thiobarbituric acid reactive substances (TBARS) concentrations, also suggesting that physical activity can be used as a therapeutic and preventive modality to mitigate degenerative processes produced due to the oxidative stress associated with age [17]. Although some information exists on the separate effects of aerobic and resistance training on circulating oxidative stress markers and HSPs concentration, there is insufficient knowledge concerning the effects of concurrent aerobicstrength exercise training on these biomarkers in older subjects for practitioners to be able to prescribe such exercise in this population. Therefore, the main purpose of this study was to examine the effects of 14 weeks of participation in a progressive concurrent training program on concentrations of HSP70 and oxidative stress markers in elderly inactive men.
2. Materials and methods 2.1. Study population Twenty four healthy elderly men, aged 55—80 years, volunteered to participate in this study. Subjects attended an information and familiarization session in which all details of the experimental procedures and potential risks were explained and only people were entered into the study after informed written consent had been obtained. Subjects with history of cardiovascular or metabolic disease, neurological disease, pulmonary disease, a physically active lifestyle, and those who used any drug medication or were smokers, were excluded from the study. Subjects were matched as well as possible according to the age and anthropometric values, then randomly allocated into either training (n = 12) and control (n = 12) groups. All subjects were asked to refrain from any exercise during the study. Moreover, to determine if both groups had similar diets, all subjects completed a validated
3 food intake questionnaire and recorded a 24-hours food in before and at the end of the study. All procedures and the investigation were approved by the committee on use of Human research subjects at Regional Research Ethics of the Mahabad Branch Islamic Azad University of Iran.
2.2. Anthropometric, performance and physiological measurements At the baseline and end of training protocol, measurements were taken for height and weight, body mass index (BMI), waist to hip ratio (WHR), and body fat percentage. To minimize technical error all measurements were taken by the same trained technician. Height and weight were measured whilst subjects were minimally clothed without shoes; subjects’ height recorded to the nearest 0.1 cm and weight (nearest 0.1 kg) using a stadiometer and digital scale. Body mass index (BMI) was calculated by diving body mass (kg) by height squared (m2 ). Waist circumference (WC) and hip circumference (HC) were measured at the midpoint between the lower border of the rib cage and the iliac crest, and at the widest part of the hip region, using a flexible tape measure subsequently. Waist to hip ratio (WHR) was calculated as waist circumference divided by that of the hip. Fat density (fat mass) was predicted from the skin-fold measurements taken on the right side of the body using a caliper (Baseline Economy ‘Slim-Guide’) at the triceps, abdominal, and super iliac sites after 10 h of fasting. Percentage body fat was then estimated by using the regression equations described by Brozek et al. [18]. Additionally, one week prior to the training, the participants in training group underwent a one-repetition maximum (1-RM) test for the five dynamic continuous exercises (chest press, leg press, leg curl, standing overhead press and ‘lat pull’) and 1RM calculated by the Brzycki equation [19].
2.3. Training protocol All participants in the training group underwent a 14 week, supervised progressive concurrent aerobic/resistance exercise program on three non-consecutive days of the week. The control group subjects were just asked to maintain their previous lifestyle and not to perform any unusual strenuous activity. Each exercise training session was done in the evening and included 45—50 min accompanied by 10 min of warm-up and 10 min of cool-down activities, which involved slow walking, calisthenics and stretching exercises. The training program began with 20 min of interchanged walking/running with the intensity controlled within the individualized 50—70% predicted maximum heart rate (HRmax) (=220 − age). Every subject wore a heart rate monitor (Polar F1mt, Finland) during running so as to maintain the correct training intensity. Then, they performed resistance training in five selected exercises with the individualized workloads at 50—70% of 1RM. Each resistance training exercise was performed as three sets of 10—12 repetitions (with 90 s recovery periods between sets). All training sessions were supervised by researchers and a qualified exercise physiologist. Also, to determine new 1RMs in different exercises as training load indicators, the 1RM testing was
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performed every three weeks and the prescribed loads were adjusted accordingly.
2.4. Blood collection and laboratory assays After a 12-hour overnight fast, blood samples were collected from antecubital vein at baseline (between 7:00 and 9:00 a.m.). A further sample was similarly collected 72 h after the completion of the training program also after an overnight fast between 7:00 and 9:00 a.m. Samples were allowed to clot at room temperature for 10 min and then were centrifuged at 3000 (4 ◦ C) rpm for 15 min. After separation, serum was aliquot into sterile micro tubes and stored at −80 ◦ C until subsequent analysis. Then concentration of Hsp70 in serum was detected using a highly sensitive enzyme linked immunosorbent assay (ELISA) technique using commercially available kits (Stress Express, USA). FRAP method was used to determine the plasma total antioxidant capacity (the sensitivity of method was 0.1 Units/ml). Plasma malondialdehyde (MDA) concentrations spectrophotometrically were assayed by measurement of thiobarbituric acid reactive substances (TBARS) assay according to the procedure of Uchiyama and Mihara [20]. In addition, serum protein carbonyls were measured by a colorimetric chemical method using an assay kit from Cayman Chemicals (Cayman Chemicals, Michigan, USA).
Figure 1 The MDA concentrations measured at baseline and after 14 week of exercise training in the both group. MDA: Malondialdehyde. *Significant training x group interaction (P < 0.01).
2.5. Statistical analyses All data are presented as mean ± SD. First, data were tested for homogeneity of variance and for normal distribution using a Kolmogorov—Smirnov and Levene’s test before statistical procedures were applied. Homogeny of two groups for anthropometric or physiological parameters at the start of the study was tested with using one-way analysis of variance (ANOVA). Also independent t-test was used to compare the results of the dietary survey. Paired student’s t-tests were used to assess group differences between groups pretraining. Then to evaluate the effects of exercise training, a two-way repeated-measures ANOVA (for time and training) was employed. All statistical analyses were performed using the SPSS statistical software package (SPSS version 20.0 for Windows, SPSS Inc., Chicago, IL, USA). The significance level was set at P ≤ 0.05.
3. Results In starting the training, there was no significant difference in the antioxidant nutrient intake between groups (P > 0.05). In addition, although total antioxidant intake increased during training in the experimental group; there remained no significant difference between training and control groups (P > 0.05). Although, the average macronutrient consumption (especially carbohydrates) in the training group increased during the intervention, these changes were not statistically significant (P > 0.05) (Table 1). Table 2 shows the body composition and anthropometrics parameters of two groups pre- and post-training. These data indicate that the groups were evenly matched prior to the training intervention. Moreover, the data clearly show that
Figure 2 The PC concentrations measured at baseline and after 14 week of exercise training in the both group. PC: protein carbonyl. *Significant training x group interaction (P < 0.01).
BMI, WC, WHR, and body fat% significantly decreased after 14 weeks of exercise training in training group (P < 0.05), while these remained unchanged in control groups (P > 0.05). The data also indicate that there was a significant difference in body mass values between two groups at the end of training program (tind = 0.030). Repeated measures analysis of variance shows that there is a very significant effect (all P < 0.01) of exercise training on all measured experimental variables when compared to the control group: MDA and PC concentrations decreasing with training, whilst HSP70 and TAC increased significantly in the training group (Figs. 1—4).
4. Discussion Accumulation of the deleterious effects of oxidative stress in the cell is believed to be a major factor in the aging process. Thus reducing oxidation of key proteins through exercise or other means may attenuate the decline in age-related function and disease prevalence [3]. The induction of HSPs, as a protective mechanism, is also decreased during aging [15], hence it would be useful to be able to implement preventive and therapeutic strategies to improve HSP induction with aging. Thus, this study was designed to examine the effects
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Effect of exercise training on oxidative stress Table 1
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Antioxidant analysis of the dietary records of the training and control groups before and after the training period. Training
Control
t
P
Protein (g/d)
Before After
88.6 ± 20.56 104 ± 12.2
95.8 ± 15.27 108 ± 20.2
0.62 0.78
0.18 0.62
CHO (g/d)
Before After
259.7 ± 45.05 280.12 ± 35.3
267.8 ± 3.89 271.3 ± 41.1
0.91 0.71
0.37 0.61
Fat (g/d)
Before After
70.1 ± 13.31 87.21 ± 12.4
83.1 ± 21.2 79.8 ± 7.7
1.64 1.13
0.11 0.27
Vitamin C (mg)
Before After
61.2 ± 21.03 65.12 ± 12.8
53 ± 19.04 58.3 ± 14.2
0.91 1.02
0.37 0.06
␣-Tocopherol (mg)
Before After
5.3 ± 2.11 6.05 ± 2.02
4.4 ± 2.06 4.8 ± 1.9
0.94 1.67
0.34 0.16
Vitamin A (g)
Before After
653.5 ± 191.18 698.7 ± 121.5
582.2 ± 174.35 592.9 ± 45.9
1.02 1.02
0.10 0.12
-Carotene (g)
Before After
922.9 ± 1306 571.3 ± 987.2
1050 ± 655.9 679.6 ± 457.3
1.01 1.21
0.32 0.06
Magnesium (mg)
Before After
228 ± 61.3 234.3 ± 29.7
214.1 ± 77 261.8 ± 44.4
0.44 0.51
0.66 0.45
Iron (mg)
Before After
15 ± 2.3 16.9 ± 1.52
16.9 ± 4.22 15.6 ± 0.8
1.14 1.35
0.23 0.12
Copper (mg)
Before After
1.42 ± 0.68 1.45 ± 0.21
1.65 ± 0.66 1.22 ± .011
0.74 0.82
0.46 0.12
Zinc (mg)
Before After
28.6 ± 15.63 30.6 ± 10.1
26.8 ± 10.76 23.3 ± 9.7
0.30 1.21
0.76 0.07
Selenium (mg)
Before After
71.7 ± 18.23 78.9 ± 16.03
81.7 ± 28.01 82.3 ± 15.3
1.13 0.22
0.27 0.82
Data expressed as mean ± SD.
Table 2
Anthropometrics and body composition parameters in two group pre and post- training.
Variable
Training
Control
Pre
Post
P
Weight (kg)
80.29 ± 3.04
78.63 ± 4.11
BMI (kg m−2 ) Body fat (%) Waist (cm) Hip (cm) WHR
27.11 ± 1.27 23.44 ± 6.77 94.20 ± 3.83 100.50 ± 2.9 0.93 ± 0.03*
26.45 ± 1.46* 20.38 ± 3.99* 90.78 ± 3.37* 100.50 ± 2.94 0.94 ± 0.03*
*,**
*
tdp = 0.004 tidp = 0.030** tdp = 0.004* tdp = 0.016* tdp = 0.03* tdp = 0.789 tdp = 0.004*
Pre
Post
82.30 ± 4.84
83.11 ± 5.30
26.93 21.70 93.50 99.99 0.92
± ± ± ± ±
1.44 3.52 4.05 3.11 0.02
27.03 22.24 93.79 99.84 0.93
P
± ± ± ± ±
1.90 3.63 4.43 3.21 0.04
**
tdp = 0.081** tidp = 0.030 tdp = 0.665 tdp = 0.118 tdp = 0.293 tdp = 0.111 tdp = 0.269
BMI: body mass index; WHR: waist-to hip ratio. * Significant difference between pre and post training. ** Significant difference between training and control groups (P < 0.05).
of a concurrent exercise program on plasma oxidative stress biomarkers and HSP70 concentrations in older men. The primary findings of this study were that, concurrent training for 14 weeks attenuates oxidative stress while simultaneously enhancing total antioxidant capacity at rest in older men. Further, exercise training was able to make significant improvements in PC as a measure of protein oxidation. Consistent with our results Campbell
et al. reported that 12-month of moderate-intensity aerobic exercise decreases oxidative stress among previously sedentary older women [21]. Moreover, recently Koubaa et al. indicated that 12-week of moderate intensity interval training program significantly increased total antioxidant status (TAS) and other antioxidant enzymes activities, whilst decreasing MDA concentrations in adults’ smokers [22]. The study of Park et al. also showed that 12-weeks low-volume
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Figure 3 The HSP70 concentrations measured at baseline and after 14 week of exercise training in the both group. HSP70: heat shock protein 70. *Significant training x group interaction (P < 0.01).
Figure 4 The TAC concentrations measured at baseline and after 14 week of exercise training in the both group. TAC: Total antioxidant capacity. *Significant training x group interaction (P < 0.01).
of physical activity is an effective intervention strategy for reducing TBARS concentrations in older adults [17]. However, it has been suggested that the beneficial effects of exercise training on oxidative stress appear depends on the characteristics of training (e.g. exercise intensity and duration, type of exercise). Although the several studies it has been done on the effect of endurance and resistance exercise separately on the oxidative stress, the effects of concurrent resistance endurance training on oxidative stress have not been well studied and the data available are limited and contradictory. Azizbeigi et al., in agreement with the finding of the present study, demonstrated that concurrent training for 8-weeks significantly decreased the MDA concentration and increased TAC and other antioxidant enzymes in untrained (younger) males [23]. Likewise, another study conducted by Schaun et al. on 20 sedentary middle age men has shown that both concurrent (aerobic-strength) and aerobic training programs can improve systemic redox status and antioxidant defense [24]. While, in contrast with the findings of the study Radovanovic et al. reported the increase in oxidative stress biomarkers in male judokas oxidative following 12 weeks concurrent training [25]. The discrepancies between the present findings and that of these researches could be the subjects’ age and physical fitness difference, as well as, the difference in exercise training protocols (intensity, duration, and frequency of various exercises). As the only exercise protocol
employed in this study was concurrent aerobic/resistance training we are unable to partition the observed effects to either the endurance (aerobic) or the resistance component of the training program. Clearly, further research is required to tease this issue out. Although delineating the exact mechanisms mediating possible changes in oxidative stress after exercise training was beyond the scope of the present study, it has been proposed that through activation of signaling pathways that augment synthesis of cellular enzymatic and nonenzymatic antioxidants [26], exercise training could have beneficial effects on the natural physiological ROS levels. In this regard, recent investigation by Ceci et al. indicated that elderly trained subjects display a higher ratio of antioxidant enzyme activity and a less pronounced increase in MDA after maximal exercise compared to sedentary subjects [5]. They concluded that this may be due to better the oxidant/antioxidant balance in athletes compared with nonathletes. Similarly, it has been shown that exercise training decreased the impact of an oxidative challenge through upregulation of antioxidant enzymes, as it is known that in the trained subjects, enhanced antioxidant defenses successfully protect the organism against increased oxidative stress and oxidative stress occurs less after exhaustive acute exercise in trained individuals [14,27]. As mentioned above, our results reveal that concurrent training for 14 weeks can improve body composition in inactive elderly men. These results are consistent with several other studies [28,29] that demonstrate an improvement in body composition after various types of training programs. Kuhle et al., in a comprehensive meta-analysis, revealed that exercise training in overweight and obese older individuals could improve some anthropometric measures such as BMI and WC [30]. Furthermore, it has been reported that concurrent exercise training can improve body composition of middle-age men with a sedentary lifestyle. HSP70 expression and (blood) concentration has also been show to decrease with aging [31], and thus may increase the increased vulnerability to oxidative stress that occurs with age [32]. However, as is corroborated by the literature, it is thought that exercise training through elevated temperature, hormones, ROS, or mechanical deformation of tissues, can induce HSP70 expression [33]. To date, the effect of concurrent exercise training on HSP70 concentration in older individual has not been deeply investigated. It is noticeable, that the results of the present study showed that concurrent exercise training can increase Hsp70 concentration in older men, and that this was associated with a reduction of oxidative damage and reduced oxidative stress. This is consistent with the results from a recent study, which showed that regular exercise training produces a remarkable increase expression of HSPs, especially HSP70, in young and old animals, and this benefit appears proportional to training frequency and duration [34]. Soufi et al. observed that in old rats, long-term training induced a marked increase in HSP70 expression compared with their sedentary counterparts [16]. Also, they suggest that exercise training, by up-regulation of antioxidant enzyme activity and increases in HSP70 levels, may attenuate apoptosis rate in rat myocardium. However, in contrast with these findings it has been found that, HSP70 protein content were unaltered after training 12 weeks of aerobic exercise training in
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Effect of exercise training on oxidative stress nine older women [35]. This divergence in these results may be explained in part, by differing in research participating cohorts (human vs. animals), the differences in measured variables (serum concentration HSP70 vs. HSP expression) which have an important role in inducing the stress response, the difference in protocols implemented (duration, and type of training methods) and gender. With the data we collected, we are unable to describe the physiological processes which might link HSP induction and reduced oxidative damage with exercise training. These may include temperature increases, and the inflammatory response, as they have previously been linked to stimulating HSP70 production. Clearly though, there is scope for a more thorough investigation, in older persons, to be performed. It is worth noting that the findings reported herein cannot be generalized to specific ages groups (e.g. frail elderly), as the range of ages in our study is wide. Access to suitable participants, and the time available to conduct the project provided limitations to our research. Nevertheless, we feel confident that prescription of concurrent training can now be prescribed to older individuals for the purpose of reducing oxidative damage, and therefore attenuate some aging processes.
Disclosure of interest The authors declare that they have no competing interest.
Acknowledgments We would like to thank our participants in our study. Also, we would like to appreciate the President of Research, Islamic Azad University, Mahabad Branch, for providing financial support for the present study.
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[31] Njemini R, Bautmans I, Onyema OO, Puyvelde KV, Demanet C, Mets T. Circulating heat shock protein 70 in health, aging and disease. BMC Immunol 2011;12:24. [32] Galli RL, Bielinski DF, Szprengiel AS, Shukitt-Hale B, Joseph JA. Blueberry supplemented diet reverses age-related decline in hippocampal HSP70 neuroprotection. Neurobiol Aging 2016;27:344—50. [33] Noble EG, Shen GX. Impact of exercise and metabolic disorders on heat shock proteins and vascular inflammation. Autoimmune Dis 2012, http://dx.doi.org/10.1155/2012/836519. ID 83651913. [34] Kim JS, Lee YH, Choi D, Yi HO. Expression of heat shock proteins (HSPs) in aged skeletal muscles depends on the frequency and duration of exercise training. J Sports Sci Med 2015;14:347—53. [35] Konopka AR, Douglass MD, Kaminsky LA, Jemiolo B, Trappe TA, Trappe S, et al. Molecular adaptations to aerobic exercise training in skeletal muscle of older women. J Gerontol A Biol Sci Med Sci 2010;65(11):1201—7.
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ORIGINAL ARTICLE
Changes of stress proteins and oxidative stress indices with progressive exercise training in elderly men Changements des protéines de stress et des marqueurs du stress oxydant lors d’un entraînement progressif chez des sujets âgés de sexe masculin S. Atashak a,∗, K. Azizbeigi b, M. Ali Azarbayjani c, S.R. Stannard d, F. Dehghan e, R. Soori e a
Department of Exercise Physiology, Mahabad Branch, Islamic Azad University, Mahabad, Iran Department of Exercise Physiology, Sanandaj Branch, Islamic Azad University, Sanandaj, Iran c Department of Exercise Physiology, Central Tehran Branch, Islamic Azad University, Tehran, Iran d School of Sport and Exercise, Massey University, New Zealand e Department of Exercise Physiology, Faculty of Physical Education and Sport Sciences, University of Tehran, Tehran, Iran b
Received 8 December 2015; accepted 16 January 2017
KEYWORDS Malondyaldeyde; Heat shocks protein-70; Carbonyl protein; Total antioxidant capacity; Elderly men
∗
Summary Purpose. — Excessive generation of free radicals and oxidative stress play an important role in acceleration of the aging process and contribute many chronic diseases. Nevertheless, it has been shown that the elderly who are physically active benefit from exercise-induced adaptation in cellular antioxidant defense systems and associated increase in the generation heat shocks proteins (HSPs). Therefore, the aim of the present research was to investigate the effects of 14 weeks participation in concurrent (resistance and aerobic) training on indices of oxidative stress and HSP70 concentration in inactive elderly men.
Corresponding author. E-mail address: [email protected] (S. Atashak).
http://dx.doi.org/10.1016/j.scispo.2017.01.006 0765-1597/© 2017 Elsevier Masson SAS. All rights reserved.
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S. Atashak et al. Methods. — Twenty-four inactive elderly men voluntarily participated in this research and were assigned to either training (n = 12) or control (n = 12) groups. In the control group, participants were advised to maintain their normal lifestyle during the study, while the training group was prescribed a combination of resistance and aerobic training for 14 weeks. At baseline and after 14 weeks, venous blood samples were obtained for measurement of malondialdehyde (MDA) and carbonyl protein (PC), total antioxidant capacity (TAC) and HSP70 concentration. Results. — The results indicate very significant effects of exercise training on all measured experimental variables when compared to the control group: MDA and PC concentrations decreasing, whilst HSP70 and TAC in aged men were increased significantly after exercise training in training group (all P < 0.01). Conclusion. — This study shows conclusively that concurrent exercise training for 14 weeks decreases indices of oxidative stress in elderly men and can ameliorate age-related deficits in HSP70 and the circulating antioxidant defense system. © 2017 Elsevier Masson SAS. All rights reserved.
MOTS CLÉS Malondyaldéhyde ; Protéine HSP 70 carbonyle ; Capacité antioxydante totale ; Sujet âgé
Résumé But. — Une production excessive de radicaux libres et le stress oxydant jouent un rôle important dans l’accélération du processus de vieillissement et sont impliquées dans beaucoup de pathologies chroniques. Néanmoins, il a été démontré que les personnes âgées qui sont physiquement actives profitent des modifications induites par l’exercice dans les systèmes de défense antioxydants et l’augmentation associée des protéines du choc thermique (HSP). Le but de ce travail était donc d’étudier les effets de la participation à 14 semaines d’entraînement associé (résistance + endurance) sur les marqueurs du stress oxydatif et les concentrations de HSP70 chez des hommes âgés sédentaires. Méthodes. — Vingt-quatre hommes âgés sédentaires ont volontairement participé à cette recherche 20 et ont été affectés aux groupes entraînement (n = 12) ou contrôle (n = 12). Dans le groupe témoin, les participants ont été invités à maintenir leur mode de vie habituel au cours de l’étude, tandis que le groupe entraînement a réalisé 14 semaines d’entraînement associé (résistance + endurance). Avant le début de l’étude et à la fin des 14 semaines, des échantillons de sang veineux ont été obtenus pour la mesure du malondialdéhyde (MDA), de la protéine carbonyle (PC), de la capacité anti-oxydante totale (TAC) et de la concentration en HSP70. Résultats. — Les résultats montrent un effet marqué de l’entraînement physique sur toutes les variables mesurées en comparaison avec le groupe contrôle : les concentrations de MDA et PC diminuent, la ue HSP70 et la TAC augmentent de fac ¸on significative après l’entraînement physique dans le groupe de sujets âgés soumis à l’entraînement (p < 0,01). Conclusion. — Cette étude montre de fac ¸on concluante que 14 semaines d’entraînement combinant résistance et endurance diminuent les marqueurs de stress oxydant chez des sujets âgés de sexe masculin et peuvent ainsi contribuer à corriger les déficits en HSP70 et en système de défense antioxydant liés à l’âge. © 2017 Elsevier Masson SAS. Tous droits r´ eserv´ es.
1. Introduction Aging, as a complex multifactorial biological process, is a progressive decline in physiological function that is often accompanied by increased susceptibility to diseases leading to disability and often premature death [1]. The free radical theory of aging, involving the accumulation of deleterious effects caused by reactive oxygen species (ROS) and reduced antioxidant defenses in elderly people, is the most popular theory proposed for the aging process [2]. According to this theory, oxidative stress increases with age and the ability of organism to cope with cellular damage induced by this stress decrease [3], finally causing cellular dysfunction and resulting in apoptosis [4]. Moreover, abundant experimental and observational studies have found that age-associated oxidative damage and reactive oxygen species (ROS) have
an important role in diverse chronic age-related diseases such as cancers, cardiovascular disease (CVD), Alzheimer’s and Parkinson’s diseases [5,6]. It seems that proteins, because they have specific functions, are one of the most obviously affected macromolecules in age-related oxidative damage [7]. An age-related increase in the protein carbonyl (PC) concentration, which has been used as a marker of ROS-mediated protein oxidation, was observed in various tissues [8]. Mutlu-Türko˘ glu et al., in this regard, observed that plasma malondialdehyde (MDA) and PC levels associated with accumulation of oxidative damage were significantly higher in the plasma of elderly subjects as compared to young subjects [9]. Moreover, it has been shown that the capacity to produce heat shock proteins (HSPs) decreases with aging in normal older subjects [10]. HSPs act as molecular
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Effect of exercise training on oxidative stress chaperones that regulate the conformation and functions of a large number of cellular proteins and has been demonstrated that to protect cells against oxidative stress injury [11]. Indeed HSPS, and especially Hsp70, may be very important in normal physiological and stressful (pro-oxidative) conditions [10]. A sedentary lifestyle typical of the elderly is a key factor in the etiology and progression of age-related chronic disease [12]. However, the majority of published data suggests that regular exercise training improves the health markers and functional capacity in older people and has considerable efficacy counteracting the aging process and associated diseases [13]. It is plausible that elderly people who engaged in physical activity, besides improving of strength, endurance and subsequently functional capacity, benefit from exerciseinduced adaptation in cellular antioxidant defense [14]. In this context, it has been reported that long-term physical training increases the activity of antioxidant enzymes and HSPs expression, and therefore reduced markers of oxidative damage [15]. In this regard, while there are indications that HSP70 expression decreases with age, long-term exercise training reduced apoptosis rate and significantly increase HSP70 in the cardiac muscles of training animals compared to sedentary control animals [16]. Further, it has also been reported that 12 week low-volume of physical activity significantly reduces the thiobarbituric acid reactive substances (TBARS) concentrations, also suggesting that physical activity can be used as a therapeutic and preventive modality to mitigate degenerative processes produced due to the oxidative stress associated with age [17]. Although some information exists on the separate effects of aerobic and resistance training on circulating oxidative stress markers and HSPs concentration, there is insufficient knowledge concerning the effects of concurrent aerobicstrength exercise training on these biomarkers in older subjects for practitioners to be able to prescribe such exercise in this population. Therefore, the main purpose of this study was to examine the effects of 14 weeks of participation in a progressive concurrent training program on concentrations of HSP70 and oxidative stress markers in elderly inactive men.
2. Materials and methods 2.1. Study population Twenty four healthy elderly men, aged 55—80 years, volunteered to participate in this study. Subjects attended an information and familiarization session in which all details of the experimental procedures and potential risks were explained and only people were entered into the study after informed written consent had been obtained. Subjects with history of cardiovascular or metabolic disease, neurological disease, pulmonary disease, a physically active lifestyle, and those who used any drug medication or were smokers, were excluded from the study. Subjects were matched as well as possible according to the age and anthropometric values, then randomly allocated into either training (n = 12) and control (n = 12) groups. All subjects were asked to refrain from any exercise during the study. Moreover, to determine if both groups had similar diets, all subjects completed a validated
3 food intake questionnaire and recorded a 24-hours food in before and at the end of the study. All procedures and the investigation were approved by the committee on use of Human research subjects at Regional Research Ethics of the Mahabad Branch Islamic Azad University of Iran.
2.2. Anthropometric, performance and physiological measurements At the baseline and end of training protocol, measurements were taken for height and weight, body mass index (BMI), waist to hip ratio (WHR), and body fat percentage. To minimize technical error all measurements were taken by the same trained technician. Height and weight were measured whilst subjects were minimally clothed without shoes; subjects’ height recorded to the nearest 0.1 cm and weight (nearest 0.1 kg) using a stadiometer and digital scale. Body mass index (BMI) was calculated by diving body mass (kg) by height squared (m2 ). Waist circumference (WC) and hip circumference (HC) were measured at the midpoint between the lower border of the rib cage and the iliac crest, and at the widest part of the hip region, using a flexible tape measure subsequently. Waist to hip ratio (WHR) was calculated as waist circumference divided by that of the hip. Fat density (fat mass) was predicted from the skin-fold measurements taken on the right side of the body using a caliper (Baseline Economy ‘Slim-Guide’) at the triceps, abdominal, and super iliac sites after 10 h of fasting. Percentage body fat was then estimated by using the regression equations described by Brozek et al. [18]. Additionally, one week prior to the training, the participants in training group underwent a one-repetition maximum (1-RM) test for the five dynamic continuous exercises (chest press, leg press, leg curl, standing overhead press and ‘lat pull’) and 1RM calculated by the Brzycki equation [19].
2.3. Training protocol All participants in the training group underwent a 14 week, supervised progressive concurrent aerobic/resistance exercise program on three non-consecutive days of the week. The control group subjects were just asked to maintain their previous lifestyle and not to perform any unusual strenuous activity. Each exercise training session was done in the evening and included 45—50 min accompanied by 10 min of warm-up and 10 min of cool-down activities, which involved slow walking, calisthenics and stretching exercises. The training program began with 20 min of interchanged walking/running with the intensity controlled within the individualized 50—70% predicted maximum heart rate (HRmax) (=220 − age). Every subject wore a heart rate monitor (Polar F1mt, Finland) during running so as to maintain the correct training intensity. Then, they performed resistance training in five selected exercises with the individualized workloads at 50—70% of 1RM. Each resistance training exercise was performed as three sets of 10—12 repetitions (with 90 s recovery periods between sets). All training sessions were supervised by researchers and a qualified exercise physiologist. Also, to determine new 1RMs in different exercises as training load indicators, the 1RM testing was
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performed every three weeks and the prescribed loads were adjusted accordingly.
2.4. Blood collection and laboratory assays After a 12-hour overnight fast, blood samples were collected from antecubital vein at baseline (between 7:00 and 9:00 a.m.). A further sample was similarly collected 72 h after the completion of the training program also after an overnight fast between 7:00 and 9:00 a.m. Samples were allowed to clot at room temperature for 10 min and then were centrifuged at 3000 (4 ◦ C) rpm for 15 min. After separation, serum was aliquot into sterile micro tubes and stored at −80 ◦ C until subsequent analysis. Then concentration of Hsp70 in serum was detected using a highly sensitive enzyme linked immunosorbent assay (ELISA) technique using commercially available kits (Stress Express, USA). FRAP method was used to determine the plasma total antioxidant capacity (the sensitivity of method was 0.1 Units/ml). Plasma malondialdehyde (MDA) concentrations spectrophotometrically were assayed by measurement of thiobarbituric acid reactive substances (TBARS) assay according to the procedure of Uchiyama and Mihara [20]. In addition, serum protein carbonyls were measured by a colorimetric chemical method using an assay kit from Cayman Chemicals (Cayman Chemicals, Michigan, USA).
Figure 1 The MDA concentrations measured at baseline and after 14 week of exercise training in the both group. MDA: Malondialdehyde. *Significant training x group interaction (P < 0.01).
2.5. Statistical analyses All data are presented as mean ± SD. First, data were tested for homogeneity of variance and for normal distribution using a Kolmogorov—Smirnov and Levene’s test before statistical procedures were applied. Homogeny of two groups for anthropometric or physiological parameters at the start of the study was tested with using one-way analysis of variance (ANOVA). Also independent t-test was used to compare the results of the dietary survey. Paired student’s t-tests were used to assess group differences between groups pretraining. Then to evaluate the effects of exercise training, a two-way repeated-measures ANOVA (for time and training) was employed. All statistical analyses were performed using the SPSS statistical software package (SPSS version 20.0 for Windows, SPSS Inc., Chicago, IL, USA). The significance level was set at P ≤ 0.05.
3. Results In starting the training, there was no significant difference in the antioxidant nutrient intake between groups (P > 0.05). In addition, although total antioxidant intake increased during training in the experimental group; there remained no significant difference between training and control groups (P > 0.05). Although, the average macronutrient consumption (especially carbohydrates) in the training group increased during the intervention, these changes were not statistically significant (P > 0.05) (Table 1). Table 2 shows the body composition and anthropometrics parameters of two groups pre- and post-training. These data indicate that the groups were evenly matched prior to the training intervention. Moreover, the data clearly show that
Figure 2 The PC concentrations measured at baseline and after 14 week of exercise training in the both group. PC: protein carbonyl. *Significant training x group interaction (P < 0.01).
BMI, WC, WHR, and body fat% significantly decreased after 14 weeks of exercise training in training group (P < 0.05), while these remained unchanged in control groups (P > 0.05). The data also indicate that there was a significant difference in body mass values between two groups at the end of training program (tind = 0.030). Repeated measures analysis of variance shows that there is a very significant effect (all P < 0.01) of exercise training on all measured experimental variables when compared to the control group: MDA and PC concentrations decreasing with training, whilst HSP70 and TAC increased significantly in the training group (Figs. 1—4).
4. Discussion Accumulation of the deleterious effects of oxidative stress in the cell is believed to be a major factor in the aging process. Thus reducing oxidation of key proteins through exercise or other means may attenuate the decline in age-related function and disease prevalence [3]. The induction of HSPs, as a protective mechanism, is also decreased during aging [15], hence it would be useful to be able to implement preventive and therapeutic strategies to improve HSP induction with aging. Thus, this study was designed to examine the effects
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Effect of exercise training on oxidative stress Table 1
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Antioxidant analysis of the dietary records of the training and control groups before and after the training period. Training
Control
t
P
Protein (g/d)
Before After
88.6 ± 20.56 104 ± 12.2
95.8 ± 15.27 108 ± 20.2
0.62 0.78
0.18 0.62
CHO (g/d)
Before After
259.7 ± 45.05 280.12 ± 35.3
267.8 ± 3.89 271.3 ± 41.1
0.91 0.71
0.37 0.61
Fat (g/d)
Before After
70.1 ± 13.31 87.21 ± 12.4
83.1 ± 21.2 79.8 ± 7.7
1.64 1.13
0.11 0.27
Vitamin C (mg)
Before After
61.2 ± 21.03 65.12 ± 12.8
53 ± 19.04 58.3 ± 14.2
0.91 1.02
0.37 0.06
␣-Tocopherol (mg)
Before After
5.3 ± 2.11 6.05 ± 2.02
4.4 ± 2.06 4.8 ± 1.9
0.94 1.67
0.34 0.16
Vitamin A (g)
Before After
653.5 ± 191.18 698.7 ± 121.5
582.2 ± 174.35 592.9 ± 45.9
1.02 1.02
0.10 0.12
-Carotene (g)
Before After
922.9 ± 1306 571.3 ± 987.2
1050 ± 655.9 679.6 ± 457.3
1.01 1.21
0.32 0.06
Magnesium (mg)
Before After
228 ± 61.3 234.3 ± 29.7
214.1 ± 77 261.8 ± 44.4
0.44 0.51
0.66 0.45
Iron (mg)
Before After
15 ± 2.3 16.9 ± 1.52
16.9 ± 4.22 15.6 ± 0.8
1.14 1.35
0.23 0.12
Copper (mg)
Before After
1.42 ± 0.68 1.45 ± 0.21
1.65 ± 0.66 1.22 ± .011
0.74 0.82
0.46 0.12
Zinc (mg)
Before After
28.6 ± 15.63 30.6 ± 10.1
26.8 ± 10.76 23.3 ± 9.7
0.30 1.21
0.76 0.07
Selenium (mg)
Before After
71.7 ± 18.23 78.9 ± 16.03
81.7 ± 28.01 82.3 ± 15.3
1.13 0.22
0.27 0.82
Data expressed as mean ± SD.
Table 2
Anthropometrics and body composition parameters in two group pre and post- training.
Variable
Training
Control
Pre
Post
P
Weight (kg)
80.29 ± 3.04
78.63 ± 4.11
BMI (kg m−2 ) Body fat (%) Waist (cm) Hip (cm) WHR
27.11 ± 1.27 23.44 ± 6.77 94.20 ± 3.83 100.50 ± 2.9 0.93 ± 0.03*
26.45 ± 1.46* 20.38 ± 3.99* 90.78 ± 3.37* 100.50 ± 2.94 0.94 ± 0.03*
*,**
*
tdp = 0.004 tidp = 0.030** tdp = 0.004* tdp = 0.016* tdp = 0.03* tdp = 0.789 tdp = 0.004*
Pre
Post
82.30 ± 4.84
83.11 ± 5.30
26.93 21.70 93.50 99.99 0.92
± ± ± ± ±
1.44 3.52 4.05 3.11 0.02
27.03 22.24 93.79 99.84 0.93
P
± ± ± ± ±
1.90 3.63 4.43 3.21 0.04
**
tdp = 0.081** tidp = 0.030 tdp = 0.665 tdp = 0.118 tdp = 0.293 tdp = 0.111 tdp = 0.269
BMI: body mass index; WHR: waist-to hip ratio. * Significant difference between pre and post training. ** Significant difference between training and control groups (P < 0.05).
of a concurrent exercise program on plasma oxidative stress biomarkers and HSP70 concentrations in older men. The primary findings of this study were that, concurrent training for 14 weeks attenuates oxidative stress while simultaneously enhancing total antioxidant capacity at rest in older men. Further, exercise training was able to make significant improvements in PC as a measure of protein oxidation. Consistent with our results Campbell
et al. reported that 12-month of moderate-intensity aerobic exercise decreases oxidative stress among previously sedentary older women [21]. Moreover, recently Koubaa et al. indicated that 12-week of moderate intensity interval training program significantly increased total antioxidant status (TAS) and other antioxidant enzymes activities, whilst decreasing MDA concentrations in adults’ smokers [22]. The study of Park et al. also showed that 12-weeks low-volume
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Figure 3 The HSP70 concentrations measured at baseline and after 14 week of exercise training in the both group. HSP70: heat shock protein 70. *Significant training x group interaction (P < 0.01).
Figure 4 The TAC concentrations measured at baseline and after 14 week of exercise training in the both group. TAC: Total antioxidant capacity. *Significant training x group interaction (P < 0.01).
of physical activity is an effective intervention strategy for reducing TBARS concentrations in older adults [17]. However, it has been suggested that the beneficial effects of exercise training on oxidative stress appear depends on the characteristics of training (e.g. exercise intensity and duration, type of exercise). Although the several studies it has been done on the effect of endurance and resistance exercise separately on the oxidative stress, the effects of concurrent resistance endurance training on oxidative stress have not been well studied and the data available are limited and contradictory. Azizbeigi et al., in agreement with the finding of the present study, demonstrated that concurrent training for 8-weeks significantly decreased the MDA concentration and increased TAC and other antioxidant enzymes in untrained (younger) males [23]. Likewise, another study conducted by Schaun et al. on 20 sedentary middle age men has shown that both concurrent (aerobic-strength) and aerobic training programs can improve systemic redox status and antioxidant defense [24]. While, in contrast with the findings of the study Radovanovic et al. reported the increase in oxidative stress biomarkers in male judokas oxidative following 12 weeks concurrent training [25]. The discrepancies between the present findings and that of these researches could be the subjects’ age and physical fitness difference, as well as, the difference in exercise training protocols (intensity, duration, and frequency of various exercises). As the only exercise protocol
employed in this study was concurrent aerobic/resistance training we are unable to partition the observed effects to either the endurance (aerobic) or the resistance component of the training program. Clearly, further research is required to tease this issue out. Although delineating the exact mechanisms mediating possible changes in oxidative stress after exercise training was beyond the scope of the present study, it has been proposed that through activation of signaling pathways that augment synthesis of cellular enzymatic and nonenzymatic antioxidants [26], exercise training could have beneficial effects on the natural physiological ROS levels. In this regard, recent investigation by Ceci et al. indicated that elderly trained subjects display a higher ratio of antioxidant enzyme activity and a less pronounced increase in MDA after maximal exercise compared to sedentary subjects [5]. They concluded that this may be due to better the oxidant/antioxidant balance in athletes compared with nonathletes. Similarly, it has been shown that exercise training decreased the impact of an oxidative challenge through upregulation of antioxidant enzymes, as it is known that in the trained subjects, enhanced antioxidant defenses successfully protect the organism against increased oxidative stress and oxidative stress occurs less after exhaustive acute exercise in trained individuals [14,27]. As mentioned above, our results reveal that concurrent training for 14 weeks can improve body composition in inactive elderly men. These results are consistent with several other studies [28,29] that demonstrate an improvement in body composition after various types of training programs. Kuhle et al., in a comprehensive meta-analysis, revealed that exercise training in overweight and obese older individuals could improve some anthropometric measures such as BMI and WC [30]. Furthermore, it has been reported that concurrent exercise training can improve body composition of middle-age men with a sedentary lifestyle. HSP70 expression and (blood) concentration has also been show to decrease with aging [31], and thus may increase the increased vulnerability to oxidative stress that occurs with age [32]. However, as is corroborated by the literature, it is thought that exercise training through elevated temperature, hormones, ROS, or mechanical deformation of tissues, can induce HSP70 expression [33]. To date, the effect of concurrent exercise training on HSP70 concentration in older individual has not been deeply investigated. It is noticeable, that the results of the present study showed that concurrent exercise training can increase Hsp70 concentration in older men, and that this was associated with a reduction of oxidative damage and reduced oxidative stress. This is consistent with the results from a recent study, which showed that regular exercise training produces a remarkable increase expression of HSPs, especially HSP70, in young and old animals, and this benefit appears proportional to training frequency and duration [34]. Soufi et al. observed that in old rats, long-term training induced a marked increase in HSP70 expression compared with their sedentary counterparts [16]. Also, they suggest that exercise training, by up-regulation of antioxidant enzyme activity and increases in HSP70 levels, may attenuate apoptosis rate in rat myocardium. However, in contrast with these findings it has been found that, HSP70 protein content were unaltered after training 12 weeks of aerobic exercise training in
Please cite this article in press as: Atashak S, et al. Changes of stress proteins and oxidative stress indices with progressive exercise training in elderly men. Sci sports (2017), http://dx.doi.org/10.1016/j.scispo.2017.01.006
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Effect of exercise training on oxidative stress nine older women [35]. This divergence in these results may be explained in part, by differing in research participating cohorts (human vs. animals), the differences in measured variables (serum concentration HSP70 vs. HSP expression) which have an important role in inducing the stress response, the difference in protocols implemented (duration, and type of training methods) and gender. With the data we collected, we are unable to describe the physiological processes which might link HSP induction and reduced oxidative damage with exercise training. These may include temperature increases, and the inflammatory response, as they have previously been linked to stimulating HSP70 production. Clearly though, there is scope for a more thorough investigation, in older persons, to be performed. It is worth noting that the findings reported herein cannot be generalized to specific ages groups (e.g. frail elderly), as the range of ages in our study is wide. Access to suitable participants, and the time available to conduct the project provided limitations to our research. Nevertheless, we feel confident that prescription of concurrent training can now be prescribed to older individuals for the purpose of reducing oxidative damage, and therefore attenuate some aging processes.
Disclosure of interest The authors declare that they have no competing interest.
Acknowledgments We would like to thank our participants in our study. Also, we would like to appreciate the President of Research, Islamic Azad University, Mahabad Branch, for providing financial support for the present study.
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Please cite this article in press as: Atashak S, et al. Changes of stress proteins and oxidative stress indices with progressive exercise training in elderly men. Sci sports (2017), http://dx.doi.org/10.1016/j.scispo.2017.01.006