Abstract
: This study examined associations between scores of depression (DEPs), thiobarbituric acid-reactive substances (TBARS), superoxide dismutase (SOD), and catalase activity (CAT) in master athletes and untrained controls. Methods: Participants were master sprinters (MS, n= 24; 50.31 6.34 year), endurance runners (ER,n= 11; 51.35 9.12 year), untrained middle-aged (CO,n= 13; 47.21 8.61 year), and young untrained (YU,n= 15; 23.70 4.02 year). CAT, SOD, and TBARS were measured in plasma using commercial kits. DEPs were measured by the Beck Depression Inventory-II. An ANOVA, Kruskal-Wallis, Pearson's, and Spearman's correlations were applied, with a signi cance level ofp 0.05. Results: The CATs of MS and YU [760.4 U L1 170.1 U L 1 and 729.9 U L 1 186.9 U L
23.70 4.02 year). CAT, SOD, and TBARS were measured in plasma using commercial kits. DEPs were measured by the Beck Depression Inventory-II. An ANOVA, Kruskal-Wallis, Pearson's, and Spearman's correlations were applied, with a signi cance level ofp 0.05. Results: The CATs of MS and YU [760.4 U L1 170.1 U L 1 and 729.9 U L 1 186.9 U L 1] were higher than CO and ER. The SOD levels in the YU and ER [84.20 U mL 1 8.52 U mL 1 and 78.24 U mL 1 6.59 U mL 1 (p< 0.0001)] were higher than CO and MS. The TBARS in CO [11.97 nmol L 1 2.35 nmol L 1 (p< 0.0001)] was higher than in YU, MS and ER. MS had lower DEPs compared to the YU [3.60 3.66 vs.12.27 9.27 (p= 0.0002)]. A negative correlation was found between CAT and DEPs for master athletes [r = 0.3921 (p= 0.0240)] and a weak correlation [r = 0.3694 (p= 0.0344)] was found between DEPs and the CAT/TBARS ratio. Conclusions: In conclusion, the training model of master sprinters may be an effective strategy for increasing CAT and reducing DEPs. Keywords:depression; master athletes; redox balance; catalase 1. Introduction Master athletes are those over 35 years old who maintain a training routine, compete in national and/or international sports championships, and represent a distinct portion of the middle-aged and elderly population [1]. These master athletes undergo intense physical training routines (from 3 to 6 sessions per week, totaling approximately 10 h or more of weekly training). The training programs of elite masters sprinters, for example, are mainly characterized by high-intensity, low-volume sessions based mainly on anaer- obic pathways [1,2]. Additionally, the weekly training of such sprinters usually includes two sessions for speed (i.e., short sprints and long sprints for speed endurance), strength (i.e., weight lifting), power (i.e., plyometric exercises), and stretching and exibility. A few low-volume, moderate-intensity cardiovascular sessions can also be performed between high-intensity anaerobic training sessions. Otherwise, the training programs of elite mas- ter endurance athletes are based mainly on low-intensity, high-volume sessions, relying primarily on aerobic pathways. Exercise modes
(i.e., short sprints and long sprints for speed endurance), strength (i.e., weight lifting), power (i.e., plyometric exercises), and stretching and exibility. A few low-volume, moderate-intensity cardiovascular sessions can also be performed between high-intensity anaerobic training sessions. Otherwise, the training programs of elite mas- ter endurance athletes are based mainly on low-intensity, high-volume sessions, relying primarily on aerobic pathways. Exercise modes and workloads are selected individually depending on the athlete's training season [1,2]. Several studies have shown that master Int. J. Environ. Res. Public Health2023,20, 4397.
Int. J. Environ. Res. Public Health2023,20, 4397 2 of 10 athletes have better physical performance, body composition, lipid pro le, blood glucose control, and attenuated biological aging when compared to untrained peers, and have thus been referred to as a healthy aging model [3,4]. The healthy aging process of master athletes may be related to several biochemical mechanisms, including lower levels of oxidative stress and increased antioxidant de- fense [5]. A decreased antioxidant defense, on the other hand, has been identi ed as a part of the pathogenesis of depression, a multifactorial disease linked to the aging process [6]. In situations in which the antioxidant system is impaired, reactive oxygen species can damage lipids, proteins, and DNA. In addition, pro-in ammatory cytokines (IL-6, TNF- ó) increase the activity of indoleamine 2,3-dioxygenase (IDO), an enzyme involved in the synthesis of kynurenine from tryptophan. Kynurenine in turn appears to have potential neurotoxic action, since kynurenine is transformed into 3-monooxygenase (KMO), forming kynurenine into 3-hydroxykynurenine and 3-hydroxyanthranilic, precursors of quinolinic acid. This acid is considered a metabolite that leads to excitotoxicity for the central nervous system and induces oxidative stress. Thus, some studies have shown that catalase seems to block the toxicity generated by 3-hydroxykynurenine [79]. Antioxidant defense, in turn, protects cells by removing free radicals. This antioxidant system comprises different types of functional components, such as superoxide dismutase (SOD) and catalase (CAT) [10]. SOD acts as a primary cellular defense against free radicals since it catalyzes the reduction of SO to oxygen and hydrogen peroxide. CAT is an antioxi- dant enzyme present in almost all aerobic organisms. Its function is to break two molecules of hydrogen peroxide into one molecule of oxygen and two molecules of water [10,11]. In our previous studies, we have shown that master athletes have greater catalase activity than their non-athlete peers, in addition to other antioxidant enzymes such as superoxide dismutase (SOD) [3,4]. However, ndings on catalase and its relationship with the intensity of depression symptoms (DEPs) are still inconsistent. According to Tsai and Huang [12], catalase activity is increased in patients in the acute phase of
we have shown that master athletes have greater catalase activity than their non-athlete peers, in addition to other antioxidant enzymes such as superoxide dismutase (SOD) [3,4]. However, ndings on catalase and its relationship with the intensity of depression symptoms (DEPs) are still inconsistent. According to Tsai and Huang [12], catalase activity is increased in patients in the acute phase of depression. On the other hand, in a meta- analysis by Jimenez-Fernandez [13], the differences in catalase levels among depressed and non-depressed people were not signi cant. Furthermore, the substance reactive to thiobarbituric acid (TBARS) is an enzyme that is abundant in the depressive process. TBARS is the main method to quantify the end products of lipid peroxidation, being considered a pro-oxidant enzyme used to measure the oxidative stress of tissues and cells [14]. This oxidative stress is de ned as an imbalance between pro- and antioxidant molecules. To the best of our knowledge, no research has been conducted on the relationship between catalase activity, oxidative stress, and the intensity of depression symptoms in people who have followed a training regimen their entire lives, such as master runner athletes. Therefore, we aimed to analyze catalase, oxidative stress, and the intensity of depression symptoms in master athletes, their non-athlete peers, and a young control group. We hypothesized that master athletes have higher catalase activity and lower intensity of depression symptoms when compared to their non-athlete peers and the youth control group. It is also hypothesized that there is a negative correlation between catalase activity and the intensity of depression symptoms. 2. Materials and Methods 2.1. Ethical Approval The study was approved by the Human Research Ethics Committee. All procedures were carried out according to the principles of the Declaration of Helsinki (466/2012). All subjects who agreed to participate in the study provided written informed consent, which had been clearly explained before participation.
Int. J. Environ. Res. Public Health2023,20, 4397 3 of 10 2.2. Participants The total sample (n= 63) was composed of 35 elite male master athletes at regional, national, and international levels and 28 untrained individuals. The master athletes were subdivided into master sprint athletes (MS,n= 24) from the 100 m, 200 m, 400 m, and 110 m hurdles, among others, and endurance runners (ER,n= 11) from 5 km to marathons and triathletes. The control groups consisted of young untrained (UY,n= 15) and middle-aged untrained controls (CO,n= 13). The youth sample was entirely collected in Brazil. These were mostly single college students. Master athletes were recruited from participants in the Brazilian Master Athletics Championship (S¢o Bernardo do Campo, Brazil, 2018), Grandprix Del Mercosur (Montevideo, Uruguay, 2019), and World Master Indoor Athletics Championship (Torón, Poland, 2019). The inclusion criteria for master athletes were: (1) systematic training for at least 10 years; and (2) active participation in national and/or international competitions until the date of data collection. The non-athlete subjects of the control group (young and middle-aged) were recruited through pamphlets and electronic advertisements in the city of Bras½lia-DF, Brazil, and met the inclusion criteria of not being trained and being healthy. The exclusion criteria for all participants were: (1) a history of cardiometabolic diseases; (2) a history of in ammatory disease and cancer; (3) a smoker; and (4) regular drug use, including hormone replacement therapy. 2.3. General Procedures Data were collected in the laboratory between 7 and 9 am, and all volunteers had not exercised in the previous 12 h and had fasted for at least 8 h. The collection protocol consisted of (a) anamnesis, to collect data referring to the health history and history of training and/or physical activity; and (b) an assessment of the intensity of depression symptoms, for which data were collected using the Beck Depression Inventory-II (BDI-II). The instrument has 21 items, and for each of them, there are four response statements, among which the subject chooses the most applicable to describe how she has been feeling in the last two weeks, including the test date [15]. These
an assessment of the intensity of depression symptoms, for which data were collected using the Beck Depression Inventory-II (BDI-II). The instrument has 21 items, and for each of them, there are four response statements, among which the subject chooses the most applicable to describe how she has been feeling in the last two weeks, including the test date [15]. These items refer to levels of intensity of depression symptoms, and the total score is the result of the sum of the individual items, reaching a maximum of 63 points. The nal score is classi ed into minimal, mild, moderate, and severe levels, thus indicating the intensity of depression. The questionnaires were administered one day before the competitions at the athletes 0 accommodations, which were usually close to the competition venue. The same researcher performed all of these assessments; and (c) collection of venous blood from the antecubital vein using a 4 mL vacutainer (with EDTA), with blood gradient centrifugation (Sirius 4000, Sieger, Brazil) for 15 min at 3800 rpm for plasma and serum isolation, and storage in a freezer ( 80 C) for further plasma analysis of catalase, superoxide dismutase (SOD), and TBARS. 2.4. Antioxidant Parameters The three antioxidant parameters used in this study were measured using commercial kits and following the manufacturer 0 s protocol. The SOD activity was measured using the SOD assay kit (Sigma Aldrich ® , California, USA), with a nal spectrophotometric reading at 450 nm; the CAT activity was measured using the Amplex TM Red Catalase assay kit (Thermo sher Scienti c ® , California, USA), with a nal spectrophotometric reading after one minute of incubation at 560 nm. 2.5. Lipid Peroxidation (TBARS) The protocol used in the present study is adapted from Ohkawa et al. (1979). Brie y, serum samples were diluted in 320 L MiliQ H2O (1:5) and added 1 mL of trichloroacetic acid (TCA) 17.5%, pH 2.0, following the addition of 1 mL of thiobarbituric acid (TBA) 0.6%, pH 2.0. After homogenization, the samples were kept in a water bath for 30 min at 95 C. The reaction was interrupted with the
al. (1979). Brie y, serum samples were diluted in 320 L MiliQ H2O (1:5) and added 1 mL of trichloroacetic acid (TCA) 17.5%, pH 2.0, following the addition of 1 mL of thiobarbituric acid (TBA) 0.6%, pH 2.0. After homogenization, the samples were kept in a water bath for 30 min at 95 C. The reaction was interrupted with the immersion of the microtubes in ice and the addition of 1 mL of TCA 70%, pH 2.0, and another incubation for 20 min at room temperature. After centrifugation (3000 rpm for 15 min) the supernatant was removed to
Int. J. Environ. Res. Public Health2023,20, 4397 4 of 10 new microtubes and taken to spectrophotometry reading at 540 nm. The concentration of lipid peroxidation products was calculated using the molar extinction coef cient equivalent for malondialdehyde (MDA equivalent = 1.56 10 5 M 1 cm 1). 2.6. Statistical Analysis The data were analyzed for normality and homogeneity using the Shapiro-Wilk test and the Levene test, respectively. The data were expressed as mean, standard deviation ( ), minimum, 25% percentile, median, 75% percentile, and maximum. A one-way ANOVA followed by Tukeys post hoc was applied for comparisons among studied groups for age, catalase and Tbars variables. Kruskal-Wallis with Dunn's test of multiple comparisons was applied to compare the groups on depression and SOD variables. The Spearman coef cient correlation was used to verify the association between catalase activity and the intensity of depression symptoms. The signi cance level was set at 5% (p< 0.05), and all procedures were performed using GraphPad Prism (v7.0, California, USA). In order to assess the clinical importance of results, the effect size was calculated and classi ed either as small (r = 0.2 to 0.49), moderate (r = 0.5 to 0.79) or large (r > 0.8) [15]. The sample size for a priori statistical power of 80% (1 = 0.80) indicated 20 participants for a signi cance level of 5% ( = 0.05) and small effect size (f = 0.4). Thus, we chose a sample of 80 subjects (20 for each studied group) [16]. 3. Results The characterization of the sample, the intensity of depression symptoms, the CAT, the SOD level, and the TBARS are expressed in Table The intensity of depression symptoms in the YU (12.27 9.27) was higher than in the MS group (3.60 3.66;p= 0.0002) and CO (4.61 2.56;p= 0.002). The CAT of MS and YU [760.4U L 1 170.1 U L 1 and 729.9 U L 1 186.9 U L 1] were higher than CO and ER [410.3 U L 1 67.24 U L 1 and 528.8 U L1 103.2 U L 1 (p< 0.0001)]. The SOD level in the YU
(3.60 3.66;p= 0.0002) and CO (4.61 2.56;p= 0.002). The CAT of MS and YU [760.4U L 1 170.1 U L 1 and 729.9 U L 1 186.9 U L 1] were higher than CO and ER [410.3 U L 1 67.24 U L 1 and 528.8 U L1 103.2 U L 1 (p< 0.0001)]. The SOD level in the YU and ER was higher than CO and MS [84.20 U mL 1 8.52 U mL 1 and78.24 U mL 1 6.59 U mL 1 (p< 0.0001)]. The TBARS in CO [11.97 nmol L 1 2.35 nmol L 1 (p< 0.0001)] was higher than in YU, MS, andER (Table). Furthermore, a negative correlation was found between CAT and the intensity of depres- sion symptoms for the entire group of master athletes [r = 0.3921 (p= 0.0240)] (FigureInt. J. Environ. Res. Public Health 2023, 20, x FOR PEER REVIEW 5 of 10 Mean and SD. 729.9 ± 186.9 b,d 410.3 ± 67.24 760.4 ± 170.1 b,d 528.8 ± 103.2 <0.0001 Minimum 318.8 301.1 523.5 415.0 25% Percentile 592.5 347.1 595.5 443.8 Median 801.1 420.6 783.5 508.0 75% Percentile 882.7 467.4 891.4 584.2 Maximum 956.6 494.4 1108 738.2 SOD (U·mL −1 ) Mean and SD. 84.20 ± 8.52 b,c 53.26 ± 13.05 61.46 ± 11.94 78.24 ± 6.59 b,c <0.0001 Minimum 71.11 32.14 40.95 65.12 25% Percentile 76.12 43.90 48.71 74.36 Median 85.83 54.34 65.38 80.28 75% Percentile 92.05 61.88 71.27 82.94 Maximum 95.92 73.22 75.64 85.82 Tbars (nmol·L −1 ) Mean and SD. 6.29 ± 1.47 d 11.97 ± 2.35 a,c,d 7.63 ± 1.39 8.88 ± 1.17 <0.0001 Minimum 4.170 7.990 5.210 6.700 25% Percentile 4.630 9.860 6.310 8.258 Median 6.380 11.63 7.810 8.860 75% Percentile 7.990 14.33 8.790 9.930 Maximum 8.240 14.94 9.990 10.46 A one-way ANOVA followed by Tukey's post hoc was applied to compare the groups. UY: young individuals; CO: control older; MS: sprint master athletes; ER: endurance runners in age, catalase, and Tbars variables. Kruskal-Wallis with Dunn’s test of multiple comparisons was applied to com- pare the groups. UY: young individuals; CO: control older; MS: sprint master athletes;
14.94 9.990 10.46 A one-way ANOVA followed by Tukey's post hoc was applied to compare the groups. UY: young individuals; CO: control older; MS: sprint master athletes; ER: endurance runners in age, catalase, and Tbars variables. Kruskal-Wallis with Dunn’s test of multiple comparisons was applied to com- pare the groups. UY: young individuals; CO: control older; MS: sprint master athletes; ER: endur- ance running on depression and SOD variables. (a) p < 0.05 in comparison to the young individuals. (b) p < 0.05 in comparison to the control older. (c) p < 0.05 in comparison to the sprint master athletes group. (d) p < 0.05 in comparison to the endurance runners. A significance level of p < 0.05 was adopted. Furthermore, a negative correlation was found between CAT and the intensity of de- pression symptoms for the entire group of master athletes [r = −0.3921 (p = 0.0240)] (Figure 1). Figure 1. Correlation analysis among catalase and intensity of depression symptoms in master ath- letes. Figure 1. Correlation analysis among catalase and intensity of depression symptoms in master athletes.
Int. J. Environ. Res. Public Health2023,20, 4397 5 of 10 Table 1.Age, intensity of depression symptoms, catalase, SOD, and Tbars of UY, CO, MS, and ER. Variables UY CO MS ER p-Value Age (years) Mean and SD. 23.70 4.02 b,c,d 47.21 8.61 50.31 6.34 51.35 9.12 <0.0001 Minimum 19.00 35.00 38.00 35.00 25% Percentile 21.00 42.00 45.00 42.00 Median 22.00 46.00 51.00 52.50 75% Percentile 27.00 50.00 55.50 58.00 Maximum 33.00 65.00 62.00 66.00 The intensity of depression symptoms Mean and SD. 12.27 9.27 b,c 4.61 2.56 3.60 3.66 7.66 5.29 0.0002 Minimum 1.000 0.000 0.000 1.000 25% Percentile 7.000 3.500 0.5000 2.500 Median 10.00 5.000 3.000 8.000 75% Percentile 16.00 6.500 6.000 11.50 Maximum 39.00 8.00 15.00 17.00 Catalase (U L 1 ) Mean and SD. 729.9 186.9 b,d 410.3 67.24 760.4 170.1 b,d 528.8 103.2 <0.0001 Minimum 318.8 301.1 523.5 415.0 25% Percentile 592.5 347.1 595.5 443.8 Median 801.1 420.6 783.5 508.0 75% Percentile 882.7 467.4 891.4 584.2 Maximum 956.6 494.4 1108 738.2 SOD (U mL 1 ) Mean and SD. 84.20 8.52 b,c 53.26 13.05 61.46 11.94 78.24 6.59 b,c <0.0001 Minimum 71.11 32.14 40.95 65.12 25% Percentile 76.12 43.90 48.71 74.36 Median 85.83 54.34 65.38 80.28 75% Percentile 92.05 61.88 71.27 82.94 Maximum 95.92 73.22 75.64 85.82 Tbars (nmol L 1 ) Mean and SD. 6.29 1.47 d 11.97 2.35 a,c,d 7.63 1.39 8.88 1.17 <0.0001 Minimum 4.170 7.990 5.210 6.700 25% Percentile 4.630 9.860 6.310 8.258 Median 6.380 11.63 7.810 8.860 75% Percentile 7.990 14.33 8.790 9.930 Maximum 8.240 14.94 9.990 10.46 A one-way ANOVA followed by Tukey 0 s post hoc was applied to compare the groups. UY: young individuals; CO: control older; MS: sprint master athletes; ER: endurance runners in age, catalase, and Tbars variables. Kruskal- Wallis with Dunn's test of multiple comparisons was applied to compare the groups. UY: young individuals; CO: control older; MS: sprint master athletes; ER: endurance running on depression and SOD variables. (a)p< 0.05 in comparison to the young individuals. (b)p< 0.05 in comparison to the control older. (c)p< 0.05 in comparison to the sprint master athletes group.(d)p<
Description
The training model of master sprinters may effectively increase catalase and reduce depression symptoms.