Mixed Circuit Training as a Non-Pharmacological Strategy to Improve Platelet Function and Oxidative Balance in Type 2 Diabetes: Role of Purinergic Signalling

Mixed Circuit Training as a Non-Pharmacological Strategy to Improve Platelet Function and Oxidative Balance in Type 2 Diabetes: Role of Purinergic Signalling | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Mixed Circuit Training as a Non-Pharmacological Strategy to Improve Platelet Function and Oxidative Balance in Type 2 Diabetes: Role of Purinergic Signalling Lucas Macedo CHAVES, Samantha Nuncio PRESTES, André Campos DE LIMA, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7436814/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 17 Mar, 2026 Read the published version in Purinergic Signalling → Version 1 posted 11 You are reading this latest preprint version Abstract This study investigates the impact of a 16-week mixed circuit training (MCT) program on purinergic signaling and oxidative stress markers in women with type 2 diabetes mellitus (T2DM), focusing on its potential to reduce platelet aggregation and thrombogenic risk. A total of 21 women with T2DM and 23 non-diabetic controls, all sedentary and middle-aged, underwent MCT twice weekly. Biochemical, hemodynamic, and oxidative stress parameters, along with platelet ectonucleotidase activity and extracellular ATP levels, were assessed pre- and post-intervention. MCT significantly decreased ectonucleotidase diphosphohydrolase (E-NTPDase) activity for ADP hydrolysis in platelets, along with a reduction in extracellular ATP levels, indicating a modulation of purinergic signaling. Additionally, exercise enhanced antioxidant defenses, increasing glutathione-S-transferase (GST) activity and vitamin C levels, while reducing myeloperoxidase (MPO) activity, a key pro-oxidant enzyme. These changes suggest a shift toward an anti-thrombotic profile, which may help mitigate diabetes-related cardiovascular risks. Mixed circuit training emerges as a valuable non-pharmacological strategy for improving platelet function and oxidative balance in T2DM. By modulating purinergic signaling and reducing oxidative stress, MCT could play a crucial role in preventing thrombogenic complications in diabetic individuals. Type 2 diabetes Purinergic signaling Oxidative stress Platelet aggregation Mixed circuit training Non-pharmacological therapy Figures Figure 1 Figure 2 What is known about this research topic? Mixed circuit training is known to improve metabolic and hemodynamic parameters in individuals with type 2 diabetes mellitus (T2DM). Purinergic signaling and oxidative stress play key roles in platelet aggregation, which is a crucial factor in T2DM complications. What this study adds and its future implications This study shows that mixed circuit training decreases ATP and ADP levels, increases antioxidant defenses, and reduces platelet aggregation potential in women with T2DM. These findings highlight the potential of exercise in modulating thrombogenic risk, supporting its use as a non-pharmacological strategy for T2DM management. Instructions for Twitter Summary Mixed circuit training reduces platelet aggregation markers and oxidative stress in women with type 2 diabetes, promoting an anti-thrombotic profile. Exercise emerges as a key tool for managing diabetes complications! #Diabetes #ExerciseScience #Health INTRODUCTION Type 2 diabetes mellitus (T2DM) is an epidemic disease with a major impact on healthcare worldwide, leading to lower quality of life and high levels of morbidity and mortality [ 1 ]. According to the Atlas of the International Diabetes Federation, in 2021 [ 2 ], 10% of adults were living with the disease in the world, which means 537 million people, furthermore, diabetes was the cause of 6.7 million deaths in the same year. Plus, it is estimated that the number of people with T2DM will reach 643 million in 2030 and 783 million in 2045, reinforcing its global impact on deteriorating health and mortality. A hyperglycemic state resulting from insulin resistance and dysfunction of pancreatic β-cells is characteristic of T2DM [ 3 ]. The main causes of morbidity and mortality of this disease are its microvascular complications, such as nephropathy, neuropathy, retinopathy and erectile dysfunction [ 4 ], and also macrovascular complications, like atherosclerosis, vascular inflammation, vasoconstriction and thrombosis that increase the risk of coronary and cerebrovascular events [ 5 ]. Macrovascular complications derive from endothelial cell dysfunction, inadequate fibrinolysis, and elevated platelet activity [ 6 ]. In this context, purinergic signaling plays an important role in platelet activation and aggregation. In response to an initial stimulus, platelets secrete granules that contain high concentrations of ATP and ADP, which bind to P2Y and P2X receptors and increase platelets' response to stimuli such as collagen and thrombin [ 7 ]. With this in mind, a mechanism for controlling platelet activation, thrombus growth and stability are the enzymes of the purinergic system, capable of controlling circulating levels of ADP and ATP [ 8 ]. The enzyme NTPDase is able to hydrolyze ATP and ADP into AMP, while the enzyme ecto 5'-nucleotidase (E-NT5) hydrolyzes AMP, forming adenosine, a potent inhibitor of platelet activation [ 8 ]. Among these, ADP stands out as a platelet aggregating agent, and even at micromolar concentrations, this nucleotide demonstrates an important pro-aggregating action in humans [ 9 ]. In addition to the components of the purinergic system, reactive oxygen species (ROS) are also of great importance for platelet activation, both for the coagulation cascade and in the fibrinolysis process, making platelets procoagulants and aggregators [ 10 ]. These ROS are formed from physiological processes, mainly in mitochondria during the oxidative phosphorylation process, but also by specific enzymes that have their concentration regulated by antioxidant mechanisms [ 11 ]. When an imbalance between oxidizing agents and antioxidants happens, it favors the formation of ROS, characterizing oxidative stress [ 12 ]. In the context of T2DM, changes in insulin sensitivity and an increase in plasma glucose concentration expand the production of ROS [ 13 , 14 ]. Low levels of enzymatic antioxidants have been reported in patients with hyperglycemia and complications related to T2DM [ 15 ]. Plus, low levels of non-enzymatic antioxidants and high levels of oxidative damage markers have been found in patients with T2DM [ 16 ], highlighting the participation of ROS in the diabetic condition. Indeed, physical exercise can enhance overall health. Strength training alone helps to improve body composition and insulin sensitivity [ 17 ]. While aerobic exercise demonstrated a reduction in blood glucose and triglyceride levels, as well as systemic blood pressure [ 18 , 19 ]. Moreover, both aerobic and resistance training act on purinergic signaling through different mechanisms [ 20 ]. Therefore, when aerobic and anaerobic exercises are performed alternately in the same session, there is a new type of exercise, called mixed circuit training [ 21 ]. It has already been demonstrated that mixed circuit physical training improves hemodynamic and anthropometric parameters, as well as reduces glycemic levels [ 22 ]. However, there are no previous studies demonstrating its impact on the activity of purinergic signaling and oxidative stress parameters in women with T2DM. In this context, our study aims to evaluate the impact of 16 weeks of mixed circuit training on purinergic signaling and oxidative stress components, plus hemodynamic, biochemical and anthropometric parameters in women with T2DM. MATERIAL AND METHODS Participants The participants of the present study were women, sedentary, aged between 40 and 60 years old. Women diagnosed with T2DM in Diabetes Group (DG) and women without T2DM diagnosis in Control Group (CG). The female sex was chosen because women are more likely to engage in physical exercise protocols. Men were excluded to avoid heterogeneity in groups. The participants should meet minimum frequency criteria of at least 75% and not miss 4 consecutive sessions. Also, alimentary habits should be maintained and participants should not do concomitant exercises during the training protocol. The final sample consisted of 44 women (21 DG and 23 CG). The DG (n = 21) was composed of sedentary women with the diagnosis of T2DM, which is characterized as fasting glucose above 126mg/dL or random glucose above 200mg/dL with unequivocal diabetes symptoms or HbA1c above 6,5% [ 23 ] in use of hypoglycemic medications, who were submitted to mixed circuit training; and the CG (n = 23) was composed of sedentary women without T2DM diagnosis who performed the same exercise protocol. This study was approved by the Ethics Committee of the Federal University of Fronteira Sul (Protocol number 4.598.914; Clinical trial number: not applicable). Every participant read and signed the Consent to Participate declaration. Data collection was carried out at 2 different times: before the start of the physical training protocol and after 16 weeks of performing the proposed exercise. Platelets and serum were separated and frozen before biochemical analysis, as described below. Nutritional questionnaire Composed of 18 questions that aim to evaluate and qualify the diet of the person being interviewed, with the objective of this research to evaluate whether the volunteers made changes in their eating habits associated with the proposed physical exercise. The questionnaire was applied in the initial collections and in the collection after the intervention. If dietary changes were noticed during the research period, the participant would be excluded from the statistical analysis. Mixed circuit training protocol The exercise protocol proposed for the participants was a mixed circuit, combining aerobic and resistance exercises in a circuit form (strength exercises followed by an aerobic exercise). The participants attended the gym at Academia Transformação, located in Chapecó-Brazil and the exercises were conducted by a qualified professional in the field of Physical Education. The volunteers performed the mixed circuit training twice a week, on non-consecutive days, for 16 weeks, totaling 32 sessions lasting 50 minutes each. The protocol was divided into 4 training mesocycles, each consisting of 4 microcycles (weekly) of training, with undulating periodization, that is, every 4 weeks the volume and intensity of training were alternated respecting the following order: Start with high volume and low intensity with rest of 30 ± 5 seconds, after four weeks it was changed to high intensity and low volume with rest of 35 ± 5 seconds, after low intensity and high volume with rest of 30 ± 5 seconds and finally, high intensity and low volume with rest of 35 ± 5 seconds. Anthropometric and hemodynamic parameters assessment Height, body mass and waist, hip and thigh circumference were evaluated according to the recommendations of the International Society for the Advancement of Kinanthropometry (ISAK) [ 24 ]. Blood pressure measurements were performed using an Aneroide Premium sphygmomanometer with a resolution of 0-300mmHg and maximum circumference of 35 cm. The blood pressure measurements were performed after 15 minutes of relaxing state. The blood pressure measurement was performed by a health care professional properly trained. Body mass index (BMI) was calculated as body mass divided by height squared (kg/m²). BMI together with the measurement of waist circumference (WC) allowed us to deduce values ​​indicative of visceral fat, which proved to be an important predictor of glucose homeostasis and mortality [ 25 ]. Visceral adipose tissue (VAT) was predicted by the protocol by Samouda et al. (2013)[ 26 ] which uses values ​​of WC, proximal thigh circumference (PTC), age and BMI and which was validated using computed tomography. In this protocol, the TAV is determined according to the area in cm 2 by the following equation: TAV = 2.15 x waist circumference – 3.63 x proximal thigh circumference + 1.46 x age + 6.22 x BMI – 92, 713 [SE (R2) = 36.88 (0.836)]. Total muscle mass (MM) was predicted by the protocol by Heymsfield et al. (2020) [ 27 ], validated based on dual emission x-ray densitometry (Dexa). In this case, the MM was determined by the equation: MM = 0.25 x weight + 0.09 x height – 0.111 x age + 0.0005 x age 2 – 0.06 x WC + 2 x race − 4.5. [SE (R2) = 1.7 (0.89)]. While the percentage of muscle mass (%MM) was determined by the ratio MM/body weight. The percentage of body fat mass (%FM) was predicted by the protocol of Lee et al. (2017) [ 28 ], validated based on Dexa. In this protocol, %FM is determined by the equation: %FM = 50.46 + 0.07 x age – 0.26 x height + 0.27 x WC ± race [SE (R2) = 3.86 (0.65)]. Total fat mass (FM) was determined by the product of %FM and body weight. Isolation of platelets: Platelets were isolated following the method described by Pilla et al. (PILLA et al., 1996) and modified by Lunkes et al. [ 29 ]. Blood was collected in vacuum tubes with 0.126 mol/L sodium citrate and was first centrifuged at 1200 rpm for 10 minutes to remove blood cells. Afterward, the platelet-rich plasma was centrifuged at 5000 rpm for 30 min and washed twice with 3.5 mmol/L isomolar HEPES buffer for 10 minutes at 5000 rpm. Finally, platelets were suspended in 500µL in isomolar HEPES buffer 3.5mmol/L. The amount of protein was determined by the Bradford method and adjusted using Comassie Blue using bovine albumin as standard to 0.4–0.6 mg/mL. Serum isolation Blood without anticoagulant was centrifuged for 15 minutes at 3500 rpm to separate the supernatant. Determination of ATP levels To quantitatively determine ATP in serum, the commercial ATP determination Kit (Invitrogen®) was used. ATP is quantified using bioluminescence from recombinant luciferase and its substrate D-luciferin. The assay is based on the need for ATP by luciferase to produce light, which was evaluated at a wavelength of 560nm. NTPDase and Ecto-5’-nucleotidase activities determination The activities of E-NTPDase 1 and E-NT5 were determined by a colorimetric assay that measures the release of inorganic phosphate. The reaction for E-NTPDase was carried out in a medium proposed by Pilla et al. [ 30 ] composed of CaCl2 5 mmol/L, NaCl 100 mmol/l, KCl 5 mmol/l, glucose 6 mmol/l and Tris–HCl buffer 50 mmol/l, pH 7.4. For E-NT5 the system is the same except that the 5 mmol/L CaCl2 is replaced by 10 mmol/L MgCl2. 20µL of platelets (8–12µg of proteins) suspended in 3.5mmol/L HEPES were added to the medium and pre-incubated for 10 minutes at 37°C. Then, the reaction was started by adding ATP or ADP at 1.0 mmol/L to measure E-NTPDase, and adenosine monophosphate (AMP) at 2.0 mmol/L for E-NT5, and incubated at 37°C for 60 minutes. Both reactions were stopped by adding 200µL of 10% trichloroacetic acid (ATA), providing a final concentration of 5%. The release of inorganic phosphate (Pi) was measured using the method of CHAN et al. [ 31 ] using malachite green as the dye and KH2PO4 as the standard, with a spectrophotometer reading at 630 nm. Control and patients were analyzed in triplicates. The specific activity of the enzyme was expressed as nmol Pi released/min/mg of protein. Oxidative stress analysis Myeloperoxidase (MPO) activity was determined in serum, using the method of Kayyali and colleagues (1991) [ 32 ]. Enzymatic activity was evaluated by a spectrophotometer using a peroxidase coupling system to a system containing phenol, 4-aminoantipyrine and H2O2. The results express in µmol the amount of quinoneimine produced in 30 minutes measured at a wavelength of 492nm. Another enzyme evaluated was glutathione-S-transferase (GST). Its antioxidant activity was determined in serum samples by the method of Warholm and collaborators (1985) [ 33 ], counting on an adequate amount of proteins in the sample, which was determined by the Bradford method and expressed as the delta absorbance of the sample. Ascorbic acid (or vitamin C) levels were measured in serum, according to the method of Roe and Kuether (1943) [ 34 ]. In this method, dehydroascorbic acid is coupled to 2,4dinitophenylhydrazine and the resulting derivative is treated with sulfuric acid (H2SO4) to produce a new color that will be measured by a spectrophotometer evaluation at a wavelength of 520nm. Total thiols were measured in serum according to the method of Ellman (1959) [ 35 ], which can be described as the reduction of 5–5'-dithiobis acid (2-nitrobenzoic acid) measured at a wavelength 412nm. The results were expressed as µmol T-SH/mL serum. Non-protein thiols were also tested in serum using the method of Ellman (1959) [ 35 ] with some modifications. 10% ATA was added to the serum and the sample consisted of the supernatant. The reaction was read at 412nm after adding 5–5'-dithiobis acid (2-nitrobenzoic acid). Results were expressed as µmol NPSH/mL serum. Assessment of lipid profile and glycated hemoglobin (HbA1c) Total Cholesterol (TC), high-density lipoprotein (HDL) and total triglycerides (TG) tests were carried out in an outsourced laboratory (Laboratório Diagnósticos do Brasil). Serum was the sample used and the analysis method was colorimetric. Low-density lipoprotein (LDL) was calculated using the Friedewald formula[ 36 ]: LDL = TC – HDL – (TG/5). HbA1c was also assessed in an outsourced laboratory (Laboratório Diagnósticos do Brasil), using the turbidimetry method. The material used for analysis was blood collected in a tube with EDTA. Statistical analysis First, the data were submitted to the Shapiro-Wilk test to verify its normality. As the data follows a normal distribution, the difference between the means was statistically analyzed by the two-way analysis of variance (ANOVA) using the statistical program Graph Pad Prism version 8.0 RESULTS Table 1 shows the hemodynamic characteristics of the groups before and after the intervention. Regarding SBP, there was a reduction in both groups after exercise. It was also possible to observe that the DG had higher post-intervention SBP levels compared to the CG (120.24 ± 7.62mmHg vs 114.04 ± 9.23mmHg). As for DBP levels, in the post-intervention period the DG had higher values than the CG (80.9 ± 5.5mmHg vs 74.6 ± 6.5mmHg). DBP levels in the same group showed no difference before and after the intervention. Table 1: Hemodynamic Parameters in the Control Group (CG) and Diabetic Group (DG) Before (Pre) and After (Post) the Application of the Circuit Mixed Training Protocol. Time CG DG SBP (mm/Hg) Pre 124,9 ± 11,40 129,90 ±11,94 Post 114,04 ± 9,23 a 120,24 ± 7,62 a % -8,70% -7,40% DBP (mm/Hg) Pre 76,6 ± 7,5 81,6 ± 7,6 Post 74,6 ± 6,5 80,9 ± 5,5 b % -2,60% -1% Table 1: Values of Systolic Blood Pressure (SBP) and Diastolic Blood Pressure (DBP) assessed in the control group (CG) and diabetic group (DG), before the application of the Circuit Mixed Training (pre) and after 16 weeks/32 training sessions (post). Data are presented as mean and standard deviation, along with the percentage difference. Statistical analysis was performed using two-way ANOVA, considering p<0.05 (a) statistically significant difference between pre and post-training within the same group; (b) statistically significant difference between groups at the same time point (p<0.05). Table 2 presents the anthropometric characteristics by groups before and after the intervention. The variables of weight, BMI, WC, CCO, WHR (waist to height ratio), TAV, MM (kg) and MA (kg) showed no difference pre-intervention and post-intervention in DG and CG, or in the same period between groups. However, in the pre-intervention period, a difference was observed between the groups regarding the variables %MM and %MA, with a higher percentage of MM in the CG compared to the DG (25.10 ± 1.45% vs 23.93 % ± 0.86) and a higher percentage of MA in the DG compared to the CG (40.95 ± 2.88% vs 38.03% ± 3.76). Table 2: Anthropometric Parameters in the Control Group (CG) and Diabetic Group (DG) Before (Pre) and After (Post) the Application of the Circuit Mixed Training Protocol. Time CG DG Weight (Kg) Pre 74,95 ± 9,54 77,07 ± 14,53 Post 74,00 ± 10,07 75,34 ± 14,8 % -1,40% -2,20% BMI (cm²) Pre 29,93 ± 4,01 31,31 ± 5,57 Post 28,76 ± 4,18 31,05 ± 5,46 % -3,90% -0,90% WC (cm) Pre 96,74 ± 11,39 102,76 ± 10,30 Post 96,22 ± 12,25 101,14 ± 9,51 % -0,50% -1,60% TC (cm) Pre 61,52 ± 6,08 65,52 ± 5,46 Post 63,24 ± 6,46 66,48 ± 5,39 % 2,80% 1,5% WHR Pre 0,60 ± 0,08 0,66 ± 0,06 Post 0,61 ± 0,08 0,64 ± 0,06 % 1,60% -3% VAT (cm²) Pre 137,57 ± 47,95 173,17 ± 42,66 Post 133,13 ± 51,66 161,98± 37,92 % -2,90% -6,50% MM (kg) Pre 18,82 ± 2,64 18,61 ± 3,80 Post 18,36 ± 2,61 18,64 ± 3,61 % -1,60% +0,16% MM% Pre 25,10 ± 1,45 23,93 ± 0,86 b Post 24,8 ± 1,40 24,38 ± 1,20 % -1,19% +1,9% AM (kg) Pre 27,84 ± 4,43 31,83 ± 7,93 Post 27,54 ± 4,62 31,22 ± 7,53 % -1,07% -1,90% AM% Pre 38,03 ± 3,76 40,95 ± 2,88 b Post 38,14 ± 3,98 40,52 ± 2,61 % 0,28% -1,10% Table 2: Values of weight in kilograms, body mass index (BMI), waist circumference (WC), thigh circumference (TC), waist-to-height ratio (WHR), visceral adipose tissue (VAT), muscle mass in kilograms (MM), muscle mass percentage (MM%), total adipose mass in kilograms (AM), and adipose mass percentage (AM%) assessed in the control group (CG) and diabetic group (DG), before the application of Circuit Mixed Training (pre) and after 16 weeks/32 training sessions (post). Data are presented as mean and standard deviation, along with percentage differences. Statistical analysis was performed using two-way ANOVA, considering p<0.05 (a) statistically significant difference between pre and post-training within the same group; (b) statistically significant difference between groups at the same time point (p<0.05). The results of the biochemical analyses can be seen in Table 3. It was possible to notice that in the pre-intervention period, HbA1c levels were increased in the DG and within the appropriate reference values for the control group, confirming that they were not diabetic (6, 36 ± 0.67% vs 5.4 ± 0.35%). Regarding lipid profile analyses, no changes were observed when TC, LDL and HDL were evaluated in both groups after the intervention. However, in the pre-intervention period, a difference was noted in HDL levels, which were higher in the CG compared to the DG (58.22 ± 10.08mg/dL vs 48.38 ± 10.56 mg/dL). Furthermore, TG levels showed a difference in the post-intervention period, being increased in the DG compared to the CG (139.9 ± 55.6mg/dL vs 92.77 ± 34.29mg/dL). Table 3 ‒ Biochemical Parameters in the Control Group (CG) and Diabetic Group (DG) Before (Pre) and After (Post) the Application of the Circuit Mixed Training Protocol. Time CG DG HbA1c (%) Pre 5,4 ± 0,35 6,36 ± 0,67 b Post 5,5 ± 0,31 6,43 ± 0,58 % 1,80% 1,1% TC (mg/dL) Pre 208,7 ± 47,32 189,57 ± 37,82 Post 201,4 ± 44,09 190,48 ± 34,26 % -3,50% 0,60% HDL (mg/dL) Pre 58,22 ± 10,08 48,38 ± 10,56 b Post 57,52 ± 10 50,57 ± 9,3 % -1,20% +4,50% LDL (mg/dL) Pre 125,43 ± 37,58 107,84 ± 32,38 Post 124,03 ± 35,22 106,43 ± 30,23 % -1,10% -1,30% TG (mg/dL) Pre 120,3 ± 44,64 146,3 ± 54,5 Post 92,77 ± 34,29 139,9 ± 55,6 b % -22,90% -4,4% Table 3: Values of glycated hemoglobin (HbA1c), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglycerides (TG) assessed in the control group (CG) and diabetic group (DG), before the application of Circuit Mixed Training (pre) and after 16 weeks/32 training sessions (post). Data are presented as mean and standard deviation, along with percentage differences. Statistical analysis was performed using two-way ANOVA, considering p<0.05 (a) statistically significant difference between pre and post-training within the same group; (b) statistically significant difference between groups at the same time point (p<0.05). The evaluation of ectonucleotidases can be seen in Figure 1. Regarding the activity of E-NTPDase, it can be seen that in the DG, ADP hydrolysis (Figure 1A) was reduced comparing pre-training and post-training (289.2 ±68.18 nmolPi/min/mg protein vs 207.1 ±61.38 nmolPi/min/mg protein). For this same parameter, a decrease in enzyme activity was also noted in CG pre and post-intervention (307.6 ±49.53 nmolPi/min/mg protein vs 172 ±32.73 nmolPi/min/mg protein). However, no changes were observed in E-NTPDase activity when the substrate was ATP (Figure 2B) or in E-NT5 activity (Figure 1C). Regarding the concentration of extracellular ATP (Figure 1D), before the physical training protocol, levels in the DG were significantly higher than in the CG (581.4 ±146.2 vs 421.5 ±83.29; P<0.0001 ). Furthermore, after 16 weeks of mixed circuit exercise there was a reduction in the concentration of extracellular ATP in the DG compared to pre-intervention (581.4 ±146.2 vs 487.4 ±84.16; p<0.05). No changes in CG were observed after physical exercise. About the oxidative stress parameters evaluated, there were favorable changes post-intervention with mixed circuit physical training (Figure 2). An increase in the antioxidant defenses GST and vitamin C was noted, at the same time as a decrease in MPO activity, both with statistical significance. Initially, the levels of protein thiols and non-protein thiols were quantified. Figure 2A demonstrates the levels of protein thiols in the DG and CG before and after physical exercise, which did not show a difference between groups or before and after the intervention. Figure 2B demonstrates the levels of non-protein thiols in the DG and CG before and after physical exercise; there were also no differences between groups or before and after the intervention. Figure 2C shows MPO activity in the DG and CG before and after physical exercise. It can be observed that in the DG, MPO activity decreased after the intervention (2.65±0.645mU/mL vs 2.254±0.33mU/mL). Also in the CG, a decrease in MPO activity was observed post-training (2.343±0.4677mU/mL vs 1.812±0.1738mU/mL). Furthermore, when comparing the two groups post-intervention, it was possible to observe decreased MPO activity in the CG compared to the DG. Another antioxidant evaluated was GST, and its activity was represented in figure 2D. When comparing the groups after the intervention, an increase in enzyme activity was noted in the DG compared to the CG (0.2916±0.098mM vs 0.1941±0.1022mM). Regarding vitamin C levels, at the pre-intervention period the CG had higher vitamin C values compared to the DG (0.1706±0.0278mg vs 0.1492±0.0061mg). It was also possible to notice an increase in vitamin C levels when comparing the pre-intervention and post-intervention DG (0.1492±0.0061mg vs 0.1760±0.0281mg). There were no changes in CG after the physical training protocol. DISCUSSION Considering the importance of the purinergic system and oxidative stress in modulating platelet activity and the fact that physical exercise is capable of modifying its parameters in a useful way, this study investigated the effects of mixed circuit physical training on the activity of ectonucleotidases in platelets, serum ATP levels and serum oxidative stress components. Hemodynamic, anthropometric and biochemical parameters (concerning the glycemic and lipid profile) were also considered. Regarding hemodynamic parameters, it is already established in the literature that T2DM decompensation modifies blood pressure levels [ 37 ], increasing the risk of cardiovascular events. The results of this study are in agreement with the already known [ 38 – 41 ] impacts of physical exercise on hemodynamic aspects found in the literature. SBP analysis showed a significant decrease after the mixed circuit exercise protocol, especially in the diabetes group. In contrast, DBP did not change significantly after 16 weeks of training. Meanwhile, it is expected that individuals without T2DM will present an HbA1c < 6, which is in line with what was observed in our control group both pre- and post-intervention. HbA1c reflects the body's glycemic levels in the last three months, being an important tool for assessing blood glucose levels in T2DM [ 42 ]. In the pre-intervention period of our study, HbA1c had increased values ​​in the DG compared to the CG, which was already expected and reinforced that participants from the control group did not have T2DM. About the lipid profile, it is known that elevated LDL and TG levels increase the risk of cardiovascular disease [ 43 ], at the same time that T2DM also expands the risk of these events [ 44 ]. This reinforces the importance of following the therapeutic goals of TC, HDL, LDL and TG levels recommended by the American Diabetes Association (ADA) [ 45 ]. In our study, a tendency to increase TC levels in the DG was observed, which may be associated with an expansion in HDL that is useful for removing excess LDL deposited in the blood vessels. This improvement can be strongly associated with physical exercise, considering that the volunteers did not make dietary interventions, which were evaluated through questionnaires about eating habits applied before and after physical training. Concerning TG values, there was no change ​​with statistical significance after the physical training protocol. In this case, dietary modifications associated with physical exercise are strongly recommended to reduce TG rates [ 43 ]. Related to the anthropometric results, no significant reductions were observed in body weight, BMI, WC, CCO, TAV, MM and MA, after 16 weeks of mixed circuit physical training. Although, a percentage tendency in decreasing TAV can be observed in both groups, which is an important result considering the contribution of adipose tissue to systemic insulin resistance, especially through the production of free fatty acids [ 46 ]. Studies that carried out intervention for a longer period [ 47 ] or with a greater frequency of physical exercise [ 48 , 49 ], showed significant improvement in the anthropometric variables analyzed. Regarding the parameters of the purinergic system evaluated in platelets, it has been proved that nucleotides and nucleosides play an important role in thrombotic regulation. ADP is one of the main promoters of platelet aggregation, activating platelets and causing them to adhere to the initial layer, expanding the thrombus [ 50 ], this role is already well established and important antiplatelet agents act in this process, inhibiting P2Y receptors on platelets [ 51 ]. Adenosine is an inhibitor of this process through A2 receptors, increasing the intracellular concentration of cyclic AMP (cAMP) and, as a result, inhibiting platelet activation [ 52 ]. Within this context, the enzymes NTPDase and NT5 perform an important role in regulating platelet activity by modulating the presence of ADP and adenosine in the extracellular environment. The literature shows that people with T2DM have increased E-NTPDase and E-NT5 activity, probably as a compensatory mechanism for the increase in nucleotides in the extracellular environment [ 53 ]. Physical exercise directly affects purinergic signaling and consequently, the activity of enzymes that hydrolyze nucleotides. Moreover, it has been observed that, acutely, exercise increases the capacity of these enzymes, which hydrolyze ATP, ADP and AMP into adenosine [ 20 , 54 ]. Regarding regular physical activity, Martins et al [ 55 ] demonstrated that the usual modifications from metabolic syndrome that precedes T2DM were reversed, showing a decrease in the activity of E-NTPDase and E- NT5, plus, a decline in coagulability modifications. Furthermore, the thrombus microenvironment is composed of other cells such as the ones from the immune system and, mainly, endothelial cells, which also play a regulatory role in platelet activity. In this sense, it was demonstrated the importance of ectonucleotidases in endothelial cells to inhibit platelet activation and aggregation [ 56 ], making it necessary, in the future, to study how mixed circuit training affects these cells. Still on the purinergic signaling, about serum ATP levels, the concentration of this nucleotide significantly increased in the diabetes group compared to the control before the intervention was performed, reinforcing that diabetes promotes an increase in ATP, activating more P2X type receptors [ 57 ]. These receptors are capable of regulating platelet activation, both directly, via P2X1 (promoting platelet activation) and P2X7 (inhibiting platelet activation), and indirectly by activating immune system cells that promote platelet activation [ 58 ]. In this study, no change in the activity of ectonucleotidases was observed between groups before the intervention protocol. However, after the mixed circuit training exercise, a decrease in E-NTPDase activity for the hydrolysis of ADP was observed, as supported in literature. Yet, the activity of E-NTPDase for the hydrolysis of ATP and E-NT5 did not change. On the other hand, after mixed circuit training, there was a significant decrease in ATP concentration in the diabetes group, demonstrating that exercise is an important regulator of the purinergic system, considering that a decrease in ATP concentration results in lower activity of P2X receptors that exacerbate platelet activity [ 58 ]. Furthermore, the decrease in the extracellular concentration of ATP associated with a decrease in the activity of enzymes, demonstrates a balance of the components of the purinergic system, characterized by a change in the pattern of this signaling as a whole. In addition, the association of the purinergic system with oxidative stress seems to play an important role in the prothrombotic state of T2DM. Lipid peroxidation of the phospholipid membrane [ 59 ], characteristic of states with high oxidative stress, causes dysfunction of the NTPDase enzyme, reducing its activity [ 60 ]. Regarding the oxidative stress parameters evaluated, thiols are non-enzymatic antioxidants that help in the structural protection of cells. In the literature, it was observed that in T2DM there is a reduction of thiols and this is correlated with an increase in glycation products, which are markers of advancement and the presence of complications [ 61 ]. However, in our results, it was not possible to observe a statistically significant change in this parameter between groups or before and after the intervention. Another antioxidant evaluated was GST, an enzyme that acts by suppressing the formation of free radicals that accentuate oxidative stress [ 62 ]. Studies already demonstrate the increase of GST-dependent antioxidant defenses after resistance and sprint training protocols, so their levels can be associated with physical activity [ 63 , 64 ]. Furthermore, it has also been demonstrated that GST can play a role in inhibiting platelet aggregation induced by ADP [ 65 ]. Similarly, in our study, both groups increased GST activity after the mixed circuit training protocol. Vitamin C (or ascorbic acid) is an important antioxidant that eliminates ROS [ 16 ]. In the literature, it has been demonstrated that in addition to its antioxidant effect, vitamin C intensifies the formation of prostaglandin E1. This metabolite increases the action of insulin and also plays an antiplatelet action [ 66 ], which reinforces the importance of increasing levels of this compound in people with T2DM. However, when searching for information about the impact of physical exercise on vitamin C, most studies added oral vitamin C supplementation to the training protocols [ 67 – 69 ], and studies that demonstrate the effect of physical exercise alone are hard to find, reiterating the importance of the results found in our study. In this context, after 16 weeks of circuit training, the diabetes group showed a significant increase in vitamin C levels, without its exogenous supplementation. Finally, the activity of MPO was evaluated, an oxidizing enzyme that produces reactive species that attack and modify the function of healthy cells [ 70 ]. Its activity showed a significant decrease after 16 weeks of mixed circuit training, especially in the diabetes group. Also, it has already been reported in the literature that increased MPO activity is associated with higher glucose and HbA1c levels [ 71 ], and that it can interact with and activate platelets [ 72 ]. So, like ADP, it acts in favor of platelet aggregation, predisposing pro-thrombotic events, which explains its importance in the context of T2DM. CONCLUSION In conclusion, the favorable effect of mixed exercise in circuit on purinergic signaling and oxidative stress in women with T2DM may be the most important mechanism disclosed by this study. The decrease in the activity of E-NTPDase for ADP hydrolysis, associated with the decrease in serum ATP, the increase in GST activity and vitamin C levels, as well as the decreased MPO activity suggest that this type of exercise modulated elements in favor of an anti-platelet aggregation parameter. Therefore, it was also possible to observe reduced blood pressure and improved biochemical parameters in the research volunteers. In a disease with rates of morbidity and mortality strongly related to its complications whose pathophysiology is based on prothrombotic events, the establishment of a pattern that regulates platelets both by decreasing pro-aggregators (ATP, ADP and MPO) and increasing anti-aggregators, explains the importance of physical exercise in regulating the function of platelets. At last, mixed circuit physical training proved to be an important therapeutic tool in controlling important parameters of T2DM in women. Declarations Author's contribution: A.M.C, L.M.C and S.N.P conceived the idea. A.M. supervised all work, collected blood samples and performed biochemistry analysis. D.Z., A.C.L and S.N.P performed the analysis related to the ectonucleotidase activities and helped in the statistical analysis. A.M, A.C.L, L.C.M and S.N.P performed the analysis associated with the oxidative stress parameters; S.L.C and A.C.L supervised the physical exercise intervention protocol; C.A.S performed the statistical analysis and corrected the results description. A.M.C, L.C.M, A.M. and C.A.S discussed the results. All authors read an approved the final version. Competing interests: The authors declare no competing interests. Conflict of interest: It is an academic work and there is no conflict of interest. Ethical approval: Ethics Committee of the Federal University of Fronteira Sul (UFFS), protocol number 4.598.914. Human Ethics and Consent to Participate declarations: The study was conducted in accordance with the Declaration of Helsinki (1964). All described methods and protocols were approved by the Ethics Committee of the Federal University of Fronteira Sul (UFFS), protocol number 4.598.914. 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17:49:33","extension":"xml","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":175003,"visible":true,"origin":"","legend":"","description":"","filename":"9af009f18f9d434ca51e04c530fb46c11structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7436814/v1/141e040d641c38d56a6907de.xml"},{"id":93429978,"identity":"c3e334ed-198f-4ca3-bcd3-e15717cd1993","added_by":"auto","created_at":"2025-10-13 17:49:33","extension":"html","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":191901,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7436814/v1/38c1270accb6eacb80714796.html"},{"id":93430588,"identity":"ee15b780-269f-4c0a-b692-9118fa2fe4bc","added_by":"auto","created_at":"2025-10-13 17:57:33","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":150658,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEctonucleotidase Activity and Extracellular ATP Levels Before (Pre) and After (Post) the Application of Circuit Mixed Training\u003c/strong\u003e. CG: control group; DG: diabetes group. \u003cstrong\u003eA\u003c/strong\u003eE-NTPDase activity for ATP hydrolysis. \u003cstrong\u003eB\u003c/strong\u003eE-NTPDase activity for ADP hydrolysis. \u003cstrong\u003eC\u003c/strong\u003eE-NT5 activity for AMP hydrolysis. \u003cstrong\u003eD\u003c/strong\u003eSerum ATP concentration. Data are presented as mean and standard deviation. Statistical analysis was performed using two-way ANOVA, considering p\u0026lt;0.05. (a) statistically significant difference between pre and post-training within the same group; (b) statistically significant difference between groups at the same time point (p\u0026lt;0.05).\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7436814/v1/5e5697f9f2945af868787de6.jpeg"},{"id":93429970,"identity":"1157936a-eb80-44ac-a7a8-e9e1afb6fef1","added_by":"auto","created_at":"2025-10-13 17:49:33","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":47581,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eOxidative stress parameters before (pre) and after (post) the application of circuit-based mixed training.\u003c/strong\u003e CG: control group; DG: diabetes group. \u003cstrong\u003eA\u003c/strong\u003e Total thiol concentration in serum. \u003cstrong\u003eB)\u003c/strong\u003eNon-protein thiol concentration in serum. \u003cstrong\u003eC\u003c/strong\u003eMyeloperoxidase (MPO) activity. \u003cstrong\u003eD\u003c/strong\u003eGlutathione-S-transferase (GST) activity. \u003cstrong\u003eE\u003c/strong\u003eVitamin C levels. Data are presented as mean and standard deviation. Statistical analysis was performed using two-way ANOVA, with p\u0026lt;0.05 indicating (a) statistically significant differences between pre- and post-training within the same group; (b) statistically significant differences between groups at the same time point (p\u0026lt;0.05).\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7436814/v1/a1188afcde54e2631cd30490.png"},{"id":105223769,"identity":"03c7815b-0085-4488-9311-be8006e0fe1a","added_by":"auto","created_at":"2026-03-23 16:10:32","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1664959,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7436814/v1/6c0b0427-2094-47c8-bc4b-d32dfafe8bb5.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eMixed Circuit Training as a Non-Pharmacological Strategy to Improve Platelet Function and Oxidative Balance in Type 2 Diabetes: Role of Purinergic Signalling \u003c/p\u003e","fulltext":[{"header":"What is known about this research topic?","content":"\u003cp\u003eMixed circuit training is known to improve metabolic and hemodynamic parameters in individuals with type 2 diabetes mellitus (T2DM). Purinergic signaling and oxidative stress play key roles in platelet aggregation, which is a crucial factor in T2DM complications.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eWhat this study adds and its future implications\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study shows that mixed circuit training decreases ATP and ADP levels, increases antioxidant defenses, and reduces platelet aggregation potential in women with T2DM. These findings highlight the potential of exercise in modulating thrombogenic risk, supporting its use as a non-pharmacological strategy for T2DM management.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInstructions for Twitter Summary\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMixed circuit training reduces platelet aggregation markers and oxidative stress in women with type 2 diabetes, promoting an anti-thrombotic profile. Exercise emerges as a key tool for managing diabetes complications! #Diabetes #ExerciseScience #Health\u003c/p\u003e"},{"header":"INTRODUCTION","content":"\u003cp\u003eType 2 diabetes mellitus (T2DM) is an epidemic disease with a major impact on healthcare worldwide, leading to lower quality of life and high levels of morbidity and mortality [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. According to the Atlas of the International Diabetes Federation, in 2021 [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e], 10% of adults were living with the disease in the world, which means 537\u0026nbsp;million people, furthermore, diabetes was the cause of 6.7\u0026nbsp;million deaths in the same year. Plus, it is estimated that the number of people with T2DM will reach 643\u0026nbsp;million in 2030 and 783\u0026nbsp;million in 2045, reinforcing its global impact on deteriorating health and mortality.\u003c/p\u003e\u003cp\u003eA hyperglycemic state resulting from insulin resistance and dysfunction of pancreatic β-cells is characteristic of T2DM [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. The main causes of morbidity and mortality of this disease are its microvascular complications, such as nephropathy, neuropathy, retinopathy and erectile dysfunction [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], and also macrovascular complications, like atherosclerosis, vascular inflammation, vasoconstriction and thrombosis that increase the risk of coronary and cerebrovascular events [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Macrovascular complications derive from endothelial cell dysfunction, inadequate fibrinolysis, and elevated platelet activity [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn this context, purinergic signaling plays an important role in platelet activation and aggregation. In response to an initial stimulus, platelets secrete granules that contain high concentrations of ATP and ADP, which bind to P2Y and P2X receptors and increase platelets' response to stimuli such as collagen and thrombin [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. With this in mind, a mechanism for controlling platelet activation, thrombus growth and stability are the enzymes of the purinergic system, capable of controlling circulating levels of ADP and ATP [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe enzyme NTPDase is able to hydrolyze ATP and ADP into AMP, while the enzyme ecto 5'-nucleotidase (E-NT5) hydrolyzes AMP, forming adenosine, a potent inhibitor of platelet activation [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. Among these, ADP stands out as a platelet aggregating agent, and even at micromolar concentrations, this nucleotide demonstrates an important pro-aggregating action in humans [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn addition to the components of the purinergic system, reactive oxygen species (ROS) are also of great importance for platelet activation, both for the coagulation cascade and in the fibrinolysis process, making platelets procoagulants and aggregators [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. These ROS are formed from physiological processes, mainly in mitochondria during the oxidative phosphorylation process, but also by specific enzymes that have their concentration regulated by antioxidant mechanisms [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. When an imbalance between oxidizing agents and antioxidants happens, it favors the formation of ROS, characterizing oxidative stress [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn the context of T2DM, changes in insulin sensitivity and an increase in plasma glucose concentration expand the production of ROS [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Low levels of enzymatic antioxidants have been reported in patients with hyperglycemia and complications related to T2DM [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Plus, low levels of non-enzymatic antioxidants and high levels of oxidative damage markers have been found in patients with T2DM [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], highlighting the participation of ROS in the diabetic condition.\u003c/p\u003e\u003cp\u003eIndeed, physical exercise can enhance overall health. Strength training alone helps to improve body composition and insulin sensitivity [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. While aerobic exercise demonstrated a reduction in blood glucose and triglyceride levels, as well as systemic blood pressure [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Moreover, both aerobic and resistance training act on purinergic signaling through different mechanisms [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Therefore, when aerobic and anaerobic exercises are performed alternately in the same session, there is a new type of exercise, called mixed circuit training [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIt has already been demonstrated that mixed circuit physical training improves hemodynamic and anthropometric parameters, as well as reduces glycemic levels [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. However, there are no previous studies demonstrating its impact on the activity of purinergic signaling and oxidative stress parameters in women with T2DM. In this context, our study aims to evaluate the impact of 16 weeks of mixed circuit training on purinergic signaling and oxidative stress components, plus hemodynamic, biochemical and anthropometric parameters in women with T2DM.\u003c/p\u003e"},{"header":"MATERIAL AND METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eParticipants\u003c/h2\u003e\u003cp\u003eThe participants of the present study were women, sedentary, aged between 40 and 60 years old. Women diagnosed with T2DM in Diabetes Group (DG) and women without T2DM diagnosis in Control Group (CG). The female sex was chosen because women are more likely to engage in physical exercise protocols. Men were excluded to avoid heterogeneity in groups.\u003c/p\u003e\u003cp\u003eThe participants should meet minimum frequency criteria of at least 75% and not miss 4 consecutive sessions. Also, alimentary habits should be maintained and participants should not do concomitant exercises during the training protocol. The final sample consisted of 44 women (21 DG and 23 CG).\u003c/p\u003e\u003cp\u003eThe DG (n\u0026thinsp;=\u0026thinsp;21) was composed of sedentary women with the diagnosis of T2DM, which is characterized as fasting glucose above 126mg/dL or random glucose above 200mg/dL with unequivocal diabetes symptoms or HbA1c above 6,5% [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] in use of hypoglycemic medications, who were submitted to mixed circuit training; and the CG (n\u0026thinsp;=\u0026thinsp;23) was composed of sedentary women without T2DM diagnosis who performed the same exercise protocol. This study was approved by the Ethics Committee of the Federal University of Fronteira Sul (Protocol number 4.598.914; Clinical trial number: not applicable). Every participant read and signed the Consent to Participate declaration.\u003c/p\u003e\u003cp\u003eData collection was carried out at 2 different times: before the start of the physical training protocol and after 16 weeks of performing the proposed exercise. Platelets and serum were separated and frozen before biochemical analysis, as described below.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eNutritional questionnaire\u003c/h3\u003e\n\u003cp\u003eComposed of 18 questions that aim to evaluate and qualify the diet of the person being interviewed, with the objective of this research to evaluate whether the volunteers made changes in their eating habits associated with the proposed physical exercise. The questionnaire was applied in the initial collections and in the collection after the intervention. If dietary changes were noticed during the research period, the participant would be excluded from the statistical analysis.\u003c/p\u003e\n\u003ch3\u003eMixed circuit training protocol\u003c/h3\u003e\n\u003cp\u003eThe exercise protocol proposed for the participants was a mixed circuit, combining aerobic and resistance exercises in a circuit form (strength exercises followed by an aerobic exercise). The participants attended the gym at Academia Transforma\u0026ccedil;\u0026atilde;o, located in Chapec\u0026oacute;-Brazil and the exercises were conducted by a qualified professional in the field of Physical Education.\u003c/p\u003e\u003cp\u003eThe volunteers performed the mixed circuit training twice a week, on non-consecutive days, for 16 weeks, totaling 32 sessions lasting 50 minutes each. The protocol was divided into 4 training mesocycles, each consisting of 4 microcycles (weekly) of training, with undulating periodization, that is, every 4 weeks the volume and intensity of training were alternated respecting the following order: Start with high volume and low intensity with rest of 30\u0026thinsp;\u0026plusmn;\u0026thinsp;5 seconds, after four weeks it was changed to high intensity and low volume with rest of 35\u0026thinsp;\u0026plusmn;\u0026thinsp;5 seconds, after low intensity and high volume with rest of 30\u0026thinsp;\u0026plusmn;\u0026thinsp;5 seconds and finally, high intensity and low volume with rest of 35\u0026thinsp;\u0026plusmn;\u0026thinsp;5 seconds.\u003c/p\u003e\n\u003ch3\u003eAnthropometric and hemodynamic parameters assessment\u003c/h3\u003e\n\u003cp\u003eHeight, body mass and waist, hip and thigh circumference were evaluated according to the recommendations of the International Society for the Advancement of Kinanthropometry (ISAK) [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eBlood pressure measurements were performed using an Aneroide Premium sphygmomanometer with a resolution of 0-300mmHg and maximum circumference of 35 cm. The blood pressure measurements were performed after 15 minutes of relaxing state. The blood pressure measurement was performed by a health care professional properly trained.\u003c/p\u003e\u003cp\u003eBody mass index (BMI) was calculated as body mass divided by height squared (kg/m\u0026sup2;). BMI together with the measurement of waist circumference (WC) allowed us to deduce values ​​indicative of visceral fat, which proved to be an important predictor of glucose homeostasis and mortality [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eVisceral adipose tissue (VAT) was predicted by the protocol by Samouda \u003cem\u003eet al.\u003c/em\u003e (2013)[\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e] which uses values ​​of WC, proximal thigh circumference (PTC), age and BMI and which was validated using computed tomography. In this protocol, the TAV is determined according to the area in cm\u003csup\u003e2\u003c/sup\u003e by the following equation: TAV\u0026thinsp;=\u0026thinsp;2.15 x waist circumference \u0026ndash; 3.63 x proximal thigh circumference\u0026thinsp;+\u0026thinsp;1.46 x age\u0026thinsp;+\u0026thinsp;6.22 x BMI \u0026ndash; 92, 713 [SE (R2)\u0026thinsp;=\u0026thinsp;36.88 (0.836)].\u003c/p\u003e\u003cp\u003eTotal muscle mass (MM) was predicted by the protocol by Heymsfield \u003cem\u003eet al.\u003c/em\u003e (2020) [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e], validated based on dual emission x-ray densitometry (Dexa). In this case, the MM was determined by the equation: MM\u0026thinsp;=\u0026thinsp;0.25 x weight\u0026thinsp;+\u0026thinsp;0.09 x height \u0026ndash; 0.111 x age\u0026thinsp;+\u0026thinsp;0.0005 x age\u003csup\u003e2\u003c/sup\u003e \u0026ndash; 0.06 x WC\u0026thinsp;+\u0026thinsp;2 x race \u0026minus;\u0026thinsp;4.5. [SE (R2)\u0026thinsp;=\u0026thinsp;1.7 (0.89)]. While the percentage of muscle mass (%MM) was determined by the ratio MM/body weight.\u003c/p\u003e\u003cp\u003eThe percentage of body fat mass (%FM) was predicted by the protocol of Lee et al. (2017) [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e], validated based on Dexa. In this protocol, %FM is determined by the equation: %FM\u0026thinsp;=\u0026thinsp;50.46\u0026thinsp;+\u0026thinsp;0.07 x age \u0026ndash; 0.26 x height\u0026thinsp;+\u0026thinsp;0.27 x WC\u0026thinsp;\u0026plusmn;\u0026thinsp;race [SE (R2)\u0026thinsp;=\u0026thinsp;3.86 (0.65)]. Total fat mass (FM) was determined by the product of %FM and body weight.\u003c/p\u003e\n\u003ch3\u003eIsolation of platelets:\u003c/h3\u003e\n\u003cp\u003ePlatelets were isolated following the method described by Pilla et al. (PILLA et al., 1996) and modified by Lunkes et al. [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Blood was collected in vacuum tubes with 0.126 mol/L sodium citrate and was first centrifuged at 1200 rpm for 10 minutes to remove blood cells. Afterward, the platelet-rich plasma was centrifuged at 5000 rpm for 30 min and washed twice with 3.5 mmol/L isomolar HEPES buffer for 10 minutes at 5000 rpm. Finally, platelets were suspended in 500\u0026micro;L in isomolar HEPES buffer 3.5mmol/L. The amount of protein was determined by the Bradford method and adjusted using Comassie Blue using bovine albumin as standard to 0.4\u0026ndash;0.6 mg/mL.\u003c/p\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eSerum isolation\u003c/h2\u003e\u003cp\u003eBlood without anticoagulant was centrifuged for 15 minutes at 3500 rpm to separate the supernatant.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eDetermination of ATP levels\u003c/h3\u003e\n\u003cp\u003eTo quantitatively determine ATP in serum, the commercial ATP determination Kit (Invitrogen\u0026reg;) was used. ATP is quantified using bioluminescence from recombinant luciferase and its substrate D-luciferin. The assay is based on the need for ATP by luciferase to produce light, which was evaluated at a wavelength of 560nm.\u003c/p\u003e\n\u003ch3\u003eNTPDase and Ecto-5’-nucleotidase activities determination\u003c/h3\u003e\n\u003cp\u003eThe activities of E-NTPDase 1 and E-NT5 were determined by a colorimetric assay that measures the release of inorganic phosphate. The reaction for E-NTPDase was carried out in a medium proposed by Pilla et al. [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e] composed of CaCl2 5 mmol/L, NaCl 100 mmol/l, KCl 5 mmol/l, glucose 6 mmol/l and Tris\u0026ndash;HCl buffer 50 mmol/l, pH 7.4. For E-NT5 the system is the same except that the 5 mmol/L CaCl2 is replaced by 10 mmol/L MgCl2. 20\u0026micro;L of platelets (8\u0026ndash;12\u0026micro;g of proteins) suspended in 3.5mmol/L HEPES were added to the medium and pre-incubated for 10 minutes at 37\u0026deg;C. Then, the reaction was started by adding ATP or ADP at 1.0 mmol/L to measure E-NTPDase, and adenosine monophosphate (AMP) at 2.0 mmol/L for E-NT5, and incubated at 37\u0026deg;C for 60 minutes. Both reactions were stopped by adding 200\u0026micro;L of 10% trichloroacetic acid (ATA), providing a final concentration of 5%. The release of inorganic phosphate (Pi) was measured using the method of CHAN et al. [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] using malachite green as the dye and KH2PO4 as the standard, with a spectrophotometer reading at 630 nm. Control and patients were analyzed in triplicates. The specific activity of the enzyme was expressed as nmol Pi released/min/mg of protein.\u003c/p\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eOxidative stress analysis\u003c/h2\u003e\u003cp\u003eMyeloperoxidase (MPO) activity was determined in serum, using the method of Kayyali and colleagues (1991) [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Enzymatic activity was evaluated by a spectrophotometer using a peroxidase coupling system to a system containing phenol, 4-aminoantipyrine and H2O2. The results express in \u0026micro;mol the amount of quinoneimine produced in 30 minutes measured at a wavelength of 492nm.\u003c/p\u003e\u003cp\u003eAnother enzyme evaluated was glutathione-S-transferase (GST). Its antioxidant activity was determined in serum samples by the method of Warholm and collaborators (1985) [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e], counting on an adequate amount of proteins in the sample, which was determined by the Bradford method and expressed as the delta absorbance of the sample.\u003c/p\u003e\u003cp\u003eAscorbic acid (or vitamin C) levels were measured in serum, according to the method of Roe and Kuether (1943) [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. In this method, dehydroascorbic acid is coupled to 2,4dinitophenylhydrazine and the resulting derivative is treated with sulfuric acid (H2SO4) to produce a new color that will be measured by a spectrophotometer evaluation at a wavelength of 520nm.\u003c/p\u003e\u003cp\u003eTotal thiols were measured in serum according to the method of Ellman (1959) [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e], which can be described as the reduction of 5\u0026ndash;5'-dithiobis acid (2-nitrobenzoic acid) measured at a wavelength 412nm. The results were expressed as \u0026micro;mol T-SH/mL serum. Non-protein thiols were also tested in serum using the method of Ellman (1959) [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e] with some modifications. 10% ATA was added to the serum and the sample consisted of the supernatant. The reaction was read at 412nm after adding 5\u0026ndash;5'-dithiobis acid (2-nitrobenzoic acid). Results were expressed as \u0026micro;mol NPSH/mL serum.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eAssessment of lipid profile and glycated hemoglobin (HbA1c)\u003c/h2\u003e\u003cp\u003eTotal Cholesterol (TC), high-density lipoprotein (HDL) and total triglycerides (TG) tests were carried out in an outsourced laboratory (Laborat\u0026oacute;rio Diagn\u0026oacute;sticos do Brasil). Serum was the sample used and the analysis method was colorimetric. Low-density lipoprotein (LDL) was calculated using the Friedewald formula[\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]: LDL\u0026thinsp;=\u0026thinsp;TC \u0026ndash; HDL \u0026ndash; (TG/5).\u003c/p\u003e\u003cp\u003eHbA1c was also assessed in an outsourced laboratory (Laborat\u0026oacute;rio Diagn\u0026oacute;sticos do Brasil), using the turbidimetry method. The material used for analysis was blood collected in a tube with EDTA.\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eFirst, the data were submitted to the Shapiro-Wilk test to verify its normality. As the data follows a normal distribution, the difference between the means was statistically analyzed by the two-way analysis of variance (ANOVA) using the statistical program Graph Pad Prism version 8.0\u003c/p\u003e\u003c/div\u003e"},{"header":"RESULTS","content":"\u003cp\u003eTable 1 shows the hemodynamic characteristics of the groups before and after the intervention. Regarding SBP, there was a reduction in both groups after exercise. It was also possible to observe that the DG had higher post-intervention SBP levels compared to the CG (120.24 \u0026plusmn; 7.62mmHg vs 114.04 \u0026plusmn; 9.23mmHg). As for DBP levels, in the post-intervention period the DG had higher values than the CG (80.9 \u0026plusmn; 5.5mmHg vs 74.6 \u0026plusmn; 6.5mmHg). DBP levels in the same group showed no difference before and after the intervention.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1:\u0026nbsp;\u003c/strong\u003eHemodynamic Parameters in the Control Group (CG) and Diabetic Group (DG) Before (Pre) and After (Post) the Application of the Circuit Mixed Training Protocol.\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTime\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eCG\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDG\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eSBP (mm/Hg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e124,9 \u0026plusmn; 11,40\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e129,90 \u0026plusmn;11,94\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e114,04 \u0026plusmn; 9,23\u003csup\u003ea\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e120,24 \u0026plusmn; 7,62\u003csup\u003ea\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-8,70%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-7,40%\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDBP (mm/Hg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e76,6 \u0026plusmn; 7,5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e81,6 \u0026plusmn; 7,6\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e74,6 \u0026plusmn; 6,5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e80,9 \u0026plusmn; 5,5\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-2,60%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1%\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eTable 1: Values of Systolic Blood Pressure (SBP) and Diastolic Blood Pressure (DBP) assessed in the control group (CG) and diabetic group (DG), before the application of the Circuit Mixed Training (pre) and after 16 weeks/32 training sessions (post). Data are presented as mean and standard deviation, along with the percentage difference. Statistical analysis was performed using two-way ANOVA, considering p\u0026lt;0.05 (a) statistically significant difference between pre and post-training within the same group; (b) statistically significant difference between groups at the same time point (p\u0026lt;0.05).\u003c/p\u003e\n\u003cp\u003eTable 2 presents the anthropometric characteristics by groups before and after the intervention. The variables of weight, BMI, WC, CCO, WHR (waist to height ratio), TAV, MM (kg) and MA (kg) showed no difference pre-intervention and post-intervention in DG and CG, or in the same period between groups. However, in the pre-intervention period, a difference was observed between the groups regarding the variables %MM and %MA, with a higher percentage of MM in the CG compared to the DG (25.10 \u0026plusmn; 1.45% vs 23.93 % \u0026plusmn; 0.86) and a higher percentage of MA in the DG compared to the CG (40.95 \u0026plusmn; 2.88% vs 38.03% \u0026plusmn; 3.76).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2:\u0026nbsp;\u003c/strong\u003eAnthropometric Parameters in the Control Group (CG) and Diabetic Group (DG) Before (Pre) and After (Post) the Application of the Circuit Mixed Training Protocol.\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTime\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eCG\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDG\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eWeight (Kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e74,95 \u0026plusmn; 9,54\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e77,07 \u0026plusmn; 14,53\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e74,00 \u0026plusmn; 10,07\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e75,34 \u0026plusmn; 14,8\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,40%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-2,20%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eBMI (cm\u0026sup2;)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e29,93 \u0026plusmn; 4,01\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e31,31 \u0026plusmn; 5,57\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e28,76 \u0026plusmn; 4,18\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e31,05 \u0026plusmn; 5,46\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-3,90%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,90%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eWC (cm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e96,74 \u0026plusmn; 11,39\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e102,76 \u0026plusmn; 10,30\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e96,22 \u0026plusmn; 12,25\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e101,14 \u0026plusmn; 9,51\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-0,50%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,60%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTC (cm)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e61,52 \u0026plusmn; 6,08\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e65,52 \u0026plusmn; 5,46\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e63,24 \u0026plusmn; 6,46\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e66,48 \u0026plusmn; 5,39\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e2,80%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,5%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eWHR\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,60 \u0026plusmn; 0,08\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,66 \u0026plusmn; 0,06\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,61 \u0026plusmn; 0,08\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,64 \u0026plusmn; 0,06\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,60%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-3%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eVAT (cm\u0026sup2;)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e137,57 \u0026plusmn; 47,95\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e173,17 \u0026plusmn; 42,66\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e133,13 \u0026plusmn; 51,66\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e161,98\u0026plusmn; 37,92\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-2,90%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-6,50%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMM (kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e18,82 \u0026plusmn; 2,64\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e18,61 \u0026plusmn; 3,80\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e18,36 \u0026plusmn; 2,61\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e18,64 \u0026plusmn; 3,61\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,60%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e+0,16%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eMM%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e25,10 \u0026plusmn; 1,45\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e23,93 \u0026plusmn; 0,86\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e24,8 \u0026plusmn; 1,40\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e24,38 \u0026plusmn; 1,20\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,19%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e+1,9%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eAM (kg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e27,84 \u0026plusmn; 4,43\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e31,83 \u0026plusmn; 7,93\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e27,54 \u0026plusmn; 4,62\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e31,22 \u0026plusmn; 7,53\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,07%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,90%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eAM%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e38,03 \u0026plusmn; 3,76\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e40,95 \u0026plusmn; 2,88\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e38,14 \u0026plusmn; 3,98\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e40,52 \u0026plusmn; 2,61\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,28%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,10%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eTable 2: Values of weight in kilograms, body mass index (BMI), waist circumference (WC), thigh circumference (TC), waist-to-height ratio (WHR), visceral adipose tissue (VAT), muscle mass in kilograms (MM), muscle mass percentage (MM%), total adipose mass in kilograms (AM), and adipose mass percentage (AM%) assessed in the control group (CG) and diabetic group (DG), before the application of Circuit Mixed Training (pre) and after 16 weeks/32 training sessions (post). Data are presented as mean and standard deviation, along with percentage differences. Statistical analysis was performed using two-way ANOVA, considering p\u0026lt;0.05 (a) statistically significant difference between pre and post-training within the same group; (b) statistically significant difference between groups at the same time point (p\u0026lt;0.05).\u003c/p\u003e\n\u003cp\u003eThe results of the biochemical analyses can be seen in Table 3. It was possible to notice that in the pre-intervention period, HbA1c levels were increased in the DG and within the appropriate reference values for the control group, confirming that they were not diabetic (6, 36 \u0026plusmn; 0.67% vs 5.4 \u0026plusmn; 0.35%). Regarding lipid profile analyses, no changes were observed when TC, LDL and HDL were evaluated in both groups after the intervention. However, in the pre-intervention period, a difference was noted in HDL levels, which were higher in the CG compared to the DG (58.22 \u0026plusmn; 10.08mg/dL vs 48.38 \u0026plusmn; 10.56 mg/dL). Furthermore, TG levels showed a difference in the post-intervention period, being increased in the DG compared to the CG (139.9 \u0026plusmn; 55.6mg/dL vs 92.77 \u0026plusmn; 34.29mg/dL).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3\u003c/strong\u003e ‒ Biochemical Parameters in the Control Group (CG) and Diabetic Group (DG) Before (Pre) and After (Post) the Application of the Circuit Mixed Training Protocol.\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTime\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eCG\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eDG\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eHbA1c (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e5,4 \u0026plusmn; 0,35\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e6,36 \u0026plusmn; 0,67\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e5,5 \u0026plusmn; 0,31\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e6,43 \u0026plusmn; 0,58\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,80%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e1,1%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTC (mg/dL)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e208,7 \u0026plusmn; 47,32\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e189,57 \u0026plusmn; 37,82\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e201,4 \u0026plusmn; 44,09\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e190,48 \u0026plusmn; 34,26\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-3,50%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e0,60%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eHDL (mg/dL)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e58,22 \u0026plusmn; 10,08\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e48,38 \u0026plusmn; 10,56\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e57,52 \u0026plusmn; 10\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e50,57 \u0026plusmn; 9,3\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,20%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e+4,50%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eLDL (mg/dL)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e125,43 \u0026plusmn; 37,58\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e107,84 \u0026plusmn; 32,38\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e124,03 \u0026plusmn; 35,22\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e106,43 \u0026plusmn; 30,23\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,10%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-1,30%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"3\" valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003eTG (mg/dL)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePre\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e120,3 \u0026plusmn; 44,64\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e146,3 \u0026plusmn; 54,5\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003ePost\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e92,77 \u0026plusmn; 34,29\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e139,9 \u0026plusmn; 55,6\u003csup\u003eb\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-22,90%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u003cstrong\u003e-4,4%\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eTable 3: Values of glycated hemoglobin (HbA1c), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglycerides (TG) assessed in the control group (CG) and diabetic group (DG), before the application of Circuit Mixed Training (pre) and after 16 weeks/32 training sessions (post). Data are presented as mean and standard deviation, along with percentage differences. Statistical analysis was performed using two-way ANOVA, considering p\u0026lt;0.05 (a) statistically significant difference between pre and post-training within the same group; (b) statistically significant difference between groups at the same time point (p\u0026lt;0.05).\u003c/p\u003e\n\u003cp\u003eThe evaluation of ectonucleotidases can be seen in Figure 1. Regarding the activity of E-NTPDase, it can be seen that in the DG, ADP hydrolysis (Figure 1A) was reduced comparing pre-training and post-training (289.2 \u0026plusmn;68.18 nmolPi/min/mg protein vs 207.1 \u0026plusmn;61.38 nmolPi/min/mg protein). For this same parameter, a decrease in enzyme activity was also noted in CG pre and post-intervention (307.6 \u0026plusmn;49.53 nmolPi/min/mg protein vs 172 \u0026plusmn;32.73 nmolPi/min/mg protein). However, no changes were observed in E-NTPDase activity when the substrate was ATP (Figure 2B) or in E-NT5 activity (Figure 1C).\u003c/p\u003e\n\u003cp\u003eRegarding the concentration of extracellular ATP (Figure 1D), before the physical training protocol, levels in the DG were significantly higher than in the CG (581.4 \u0026plusmn;146.2 vs 421.5 \u0026plusmn;83.29; P\u0026lt;0.0001 ). Furthermore, after 16 weeks of mixed circuit exercise there was a reduction in the concentration of extracellular ATP in the DG compared to pre-intervention (581.4 \u0026plusmn;146.2 vs 487.4 \u0026plusmn;84.16; p\u0026lt;0.05). No changes in CG were observed after physical exercise.\u003c/p\u003e\n\u003cp\u003eAbout the oxidative stress parameters evaluated, there were favorable changes post-intervention with mixed circuit physical training (Figure 2). An increase in the antioxidant defenses GST and vitamin C was noted, at the same time as a decrease in MPO activity, both with statistical significance.\u003c/p\u003e\n\u003cp\u003eInitially, the levels of protein thiols and non-protein thiols were quantified. Figure 2A demonstrates the levels of protein thiols in the DG and CG before and after physical exercise, which did not show a difference between groups or before and after the intervention. Figure 2B demonstrates the levels of non-protein thiols in the DG and CG before and after physical exercise; there were also no differences between groups or before and after the intervention.\u003c/p\u003e\n\u003cp\u003eFigure 2C shows MPO activity in the DG and CG before and after physical exercise. It can be observed that in the DG, MPO activity decreased after the intervention (2.65\u0026plusmn;0.645mU/mL vs 2.254\u0026plusmn;0.33mU/mL). Also in the CG, a decrease in MPO activity was observed post-training (2.343\u0026plusmn;0.4677mU/mL vs 1.812\u0026plusmn;0.1738mU/mL). Furthermore, when comparing the two groups post-intervention, it was possible to observe decreased MPO activity in the CG compared to the DG.\u003c/p\u003e\n\u003cp\u003eAnother antioxidant evaluated was GST, and its activity was represented in figure 2D. When comparing the groups after the intervention, an increase in enzyme activity was noted in the DG compared to the CG (0.2916\u0026plusmn;0.098mM vs 0.1941\u0026plusmn;0.1022mM). Regarding vitamin C levels, at the pre-intervention period the CG had higher vitamin C values compared to the DG (0.1706\u0026plusmn;0.0278mg vs 0.1492\u0026plusmn;0.0061mg). It was also possible to notice an increase in vitamin C levels when comparing the pre-intervention and post-intervention DG (0.1492\u0026plusmn;0.0061mg vs 0.1760\u0026plusmn;0.0281mg). There were no changes in CG after the physical training protocol.\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eConsidering the importance of the purinergic system and oxidative stress in modulating platelet activity and the fact that physical exercise is capable of modifying its parameters in a useful way, this study investigated the effects of mixed circuit physical training on the activity of ectonucleotidases in platelets, serum ATP levels and serum oxidative stress components. Hemodynamic, anthropometric and biochemical parameters (concerning the glycemic and lipid profile) were also considered.\u003c/p\u003e\u003cp\u003eRegarding hemodynamic parameters, it is already established in the literature that T2DM decompensation modifies blood pressure levels [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e], increasing the risk of cardiovascular events. The results of this study are in agreement with the already known [\u003cspan additionalcitationids=\"CR39 CR40\" citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e] impacts of physical exercise on hemodynamic aspects found in the literature. SBP analysis showed a significant decrease after the mixed circuit exercise protocol, especially in the diabetes group. In contrast, DBP did not change significantly after 16 weeks of training.\u003c/p\u003e\u003cp\u003eMeanwhile, it is expected that individuals without T2DM will present an HbA1c\u0026thinsp;\u0026lt;\u0026thinsp;6, which is in line with what was observed in our control group both pre- and post-intervention. HbA1c reflects the body's glycemic levels in the last three months, being an important tool for assessing blood glucose levels in T2DM [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. In the pre-intervention period of our study, HbA1c had increased values ​​in the DG compared to the CG, which was already expected and reinforced that participants from the control group did not have T2DM.\u003c/p\u003e\u003cp\u003eAbout the lipid profile, it is known that elevated LDL and TG levels increase the risk of cardiovascular disease [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e], at the same time that T2DM also expands the risk of these events [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. This reinforces the importance of following the therapeutic goals of TC, HDL, LDL and TG levels recommended by the American Diabetes Association (ADA) [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn our study, a tendency to increase TC levels in the DG was observed, which may be associated with an expansion in HDL that is useful for removing excess LDL deposited in the blood vessels. This improvement can be strongly associated with physical exercise, considering that the volunteers did not make dietary interventions, which were evaluated through questionnaires about eating habits applied before and after physical training. Concerning TG values, there was no change ​​with statistical significance after the physical training protocol. In this case, dietary modifications associated with physical exercise are strongly recommended to reduce TG rates [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eRelated to the anthropometric results, no significant reductions were observed in body weight, BMI, WC, CCO, TAV, MM and MA, after 16 weeks of mixed circuit physical training. Although, a percentage tendency in decreasing TAV can be observed in both groups, which is an important result considering the contribution of adipose tissue to systemic insulin resistance, especially through the production of free fatty acids [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. Studies that carried out intervention for a longer period [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e] or with a greater frequency of physical exercise [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e], showed significant improvement in the anthropometric variables analyzed.\u003c/p\u003e\u003cp\u003eRegarding the parameters of the purinergic system evaluated in platelets, it has been proved that nucleotides and nucleosides play an important role in thrombotic regulation. ADP is one of the main promoters of platelet aggregation, activating platelets and causing them to adhere to the initial layer, expanding the thrombus [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e], this role is already well established and important antiplatelet agents act in this process, inhibiting P2Y receptors on platelets [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. Adenosine is an inhibitor of this process through A2 receptors, increasing the intracellular concentration of cyclic AMP (cAMP) and, as a result, inhibiting platelet activation [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e]. Within this context, the enzymes NTPDase and NT5 perform an important role in regulating platelet activity by modulating the presence of ADP and adenosine in the extracellular environment.\u003c/p\u003e\u003cp\u003eThe literature shows that people with T2DM have increased E-NTPDase and E-NT5 activity, probably as a compensatory mechanism for the increase in nucleotides in the extracellular environment [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]. Physical exercise directly affects purinergic signaling and consequently, the activity of enzymes that hydrolyze nucleotides. Moreover, it has been observed that, acutely, exercise increases the capacity of these enzymes, which hydrolyze ATP, ADP and AMP into adenosine [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. Regarding regular physical activity, Martins et al [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e] demonstrated that the usual modifications from metabolic syndrome that precedes T2DM were reversed, showing a decrease in the activity of E-NTPDase and E- NT5, plus, a decline in coagulability modifications.\u003c/p\u003e\u003cp\u003eFurthermore, the thrombus microenvironment is composed of other cells such as the ones from the immune system and, mainly, endothelial cells, which also play a regulatory role in platelet activity. In this sense, it was demonstrated the importance of ectonucleotidases in endothelial cells to inhibit platelet activation and aggregation [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e], making it necessary, in the future, to study how mixed circuit training affects these cells.\u003c/p\u003e\u003cp\u003eStill on the purinergic signaling, about serum ATP levels, the concentration of this nucleotide significantly increased in the diabetes group compared to the control before the intervention was performed, reinforcing that diabetes promotes an increase in ATP, activating more P2X type receptors [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]. These receptors are capable of regulating platelet activation, both directly, via P2X1 (promoting platelet activation) and P2X7 (inhibiting platelet activation), and indirectly by activating immune system cells that promote platelet activation [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eIn this study, no change in the activity of ectonucleotidases was observed between groups before the intervention protocol. However, after the mixed circuit training exercise, a decrease in E-NTPDase activity for the hydrolysis of ADP was observed, as supported in literature. Yet, the activity of E-NTPDase for the hydrolysis of ATP and E-NT5 did not change.\u003c/p\u003e\u003cp\u003eOn the other hand, after mixed circuit training, there was a significant decrease in ATP concentration in the diabetes group, demonstrating that exercise is an important regulator of the purinergic system, considering that a decrease in ATP concentration results in lower activity of P2X receptors that exacerbate platelet activity [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e]. Furthermore, the decrease in the extracellular concentration of ATP associated with a decrease in the activity of enzymes, demonstrates a balance of the components of the purinergic system, characterized by a change in the pattern of this signaling as a whole.\u003c/p\u003e\u003cp\u003eIn addition, the association of the purinergic system with oxidative stress seems to play an important role in the prothrombotic state of T2DM. Lipid peroxidation of the phospholipid membrane [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e], characteristic of states with high oxidative stress, causes dysfunction of the NTPDase enzyme, reducing its activity [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eRegarding the oxidative stress parameters evaluated, thiols are non-enzymatic antioxidants that help in the structural protection of cells. In the literature, it was observed that in T2DM there is a reduction of thiols and this is correlated with an increase in glycation products, which are markers of advancement and the presence of complications [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e]. However, in our results, it was not possible to observe a statistically significant change in this parameter between groups or before and after the intervention.\u003c/p\u003e\u003cp\u003eAnother antioxidant evaluated was GST, an enzyme that acts by suppressing the formation of free radicals that accentuate oxidative stress [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e]. Studies already demonstrate the increase of GST-dependent antioxidant defenses after resistance and sprint training protocols, so their levels can be associated with physical activity [\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e, \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e]. Furthermore, it has also been demonstrated that GST can play a role in inhibiting platelet aggregation induced by ADP [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e]. Similarly, in our study, both groups increased GST activity after the mixed circuit training protocol.\u003c/p\u003e\u003cp\u003eVitamin C (or ascorbic acid) is an important antioxidant that eliminates ROS [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. In the literature, it has been demonstrated that in addition to its antioxidant effect, vitamin C intensifies the formation of prostaglandin E1. This metabolite increases the action of insulin and also plays an antiplatelet action [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e], which reinforces the importance of increasing levels of this compound in people with T2DM.\u003c/p\u003e\u003cp\u003eHowever, when searching for information about the impact of physical exercise on vitamin C, most studies added oral vitamin C supplementation to the training protocols [\u003cspan additionalcitationids=\"CR68\" citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e], and studies that demonstrate the effect of physical exercise alone are hard to find, reiterating the importance of the results found in our study. In this context, after 16 weeks of circuit training, the diabetes group showed a significant increase in vitamin C levels, without its exogenous supplementation.\u003c/p\u003e\u003cp\u003eFinally, the activity of MPO was evaluated, an oxidizing enzyme that produces reactive species that attack and modify the function of healthy cells [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e]. Its activity showed a significant decrease after 16 weeks of mixed circuit training, especially in the diabetes group. Also, it has already been reported in the literature that increased MPO activity is associated with higher glucose and HbA1c levels [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e], and that it can interact with and activate platelets [\u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e72\u003c/span\u003e]. So, like ADP, it acts in favor of platelet aggregation, predisposing pro-thrombotic events, which explains its importance in the context of T2DM.\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eIn conclusion, the favorable effect of mixed exercise in circuit on purinergic signaling and oxidative stress in women with T2DM may be the most important mechanism disclosed by this study. The decrease in the activity of E-NTPDase for ADP hydrolysis, associated with the decrease in serum ATP, the increase in GST activity and vitamin C levels, as well as the decreased MPO activity suggest that this type of exercise modulated elements in favor of an anti-platelet aggregation parameter. Therefore, it was also possible to observe reduced blood pressure and improved biochemical parameters in the research volunteers.\u003c/p\u003e\u003cp\u003eIn a disease with rates of morbidity and mortality strongly related to its complications whose pathophysiology is based on prothrombotic events, the establishment of a pattern that regulates platelets both by decreasing pro-aggregators (ATP, ADP and MPO) and increasing anti-aggregators, explains the importance of physical exercise in regulating the function of platelets. At last, mixed circuit physical training proved to be an important therapeutic tool in controlling important parameters of T2DM in women.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor's contribution: \u003c/strong\u003eA.M.C, L.M.C and S.N.P conceived the idea. A.M. supervised all work, collected blood samples and performed biochemistry analysis. D.Z., A.C.L and S.N.P performed the analysis related to the ectonucleotidase activities and helped in the statistical analysis. A.M, A.C.L, L.C.M and S.N.P performed the analysis associated with the oxidative stress parameters; S.L.C and A.C.L supervised the physical exercise intervention protocol; C.A.S performed the statistical analysis and corrected the results description. A.M.C, L.C.M, A.M. and C.A.S discussed the results. All authors read an approved the final version.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests:\u003c/strong\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest:\u003c/strong\u003eIt is an academic work and there is no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval: \u003c/strong\u003eEthics Committee of the Federal University of Fronteira Sul (UFFS), protocol number 4.598.914.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHuman Ethics and Consent to Participate declarations: \u003c/strong\u003eThe study was conducted in accordance with the Declaration of Helsinki (1964). All described methods and protocols were approved by the Ethics Committee of the Federal University of Fronteira Sul (UFFS), protocol number 4.598.914. Each participant has read and signed the Consent to Participate Declaration before the beginning of the experimental protocols. \u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e Funding:\u003c/strong\u003ethis study received financial support from Federal University of Fronteira Sul \u003c/p\u003e\u003cp\u003eData is provided within the manuscript or upon request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eKhan MAB, Hashim MJ, King JK et al (2019) Epidemiology of Type 2 Diabetes \u0026ndash; Global Burden of Disease and Forecasted Trends: JEGH 10:107. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2991/jegh.k.191028.001\u003c/span\u003e\u003cspan address=\"10.2991/jegh.k.191028.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFederation ID (2019) IDF DIABETES ATLAS, 9th edn. 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Free Radic Biol Med 61:357\u0026ndash;369. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.freeradbiomed.2013.04.014\u003c/span\u003e\u003cspan address=\"10.1016/j.freeradbiomed.2013.04.014\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"purinergic-signalling","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pusi","sideBox":"Learn more about [Purinergic Signalling](http://link.springer.com/journal/11302)","snPcode":"11302","submissionUrl":"https://submission.nature.com/new-submission/11302/3","title":"Purinergic Signalling","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Type 2 diabetes, Purinergic signaling, Oxidative stress, Platelet aggregation, Mixed circuit training, Non-pharmacological therapy","lastPublishedDoi":"10.21203/rs.3.rs-7436814/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7436814/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study investigates the impact of a 16-week mixed circuit training (MCT) program on purinergic signaling and oxidative stress markers in women with type 2 diabetes mellitus (T2DM), focusing on its potential to reduce platelet aggregation and thrombogenic risk. A total of 21 women with T2DM and 23 non-diabetic controls, all sedentary and middle-aged, underwent MCT twice weekly. Biochemical, hemodynamic, and oxidative stress parameters, along with platelet ectonucleotidase activity and extracellular ATP levels, were assessed pre- and post-intervention. MCT significantly decreased ectonucleotidase diphosphohydrolase (E-NTPDase) activity for ADP hydrolysis in platelets, along with a reduction in extracellular ATP levels, indicating a modulation of purinergic signaling. Additionally, exercise enhanced antioxidant defenses, increasing glutathione-S-transferase (GST) activity and vitamin C levels, while reducing myeloperoxidase (MPO) activity, a key pro-oxidant enzyme. These changes suggest a shift toward an anti-thrombotic profile, which may help mitigate diabetes-related cardiovascular risks. Mixed circuit training emerges as a valuable non-pharmacological strategy for improving platelet function and oxidative balance in T2DM. By modulating purinergic signaling and reducing oxidative stress, MCT could play a crucial role in preventing thrombogenic complications in diabetic individuals.\u003c/p\u003e","manuscriptTitle":"Mixed Circuit Training as a Non-Pharmacological Strategy to Improve Platelet Function and Oxidative Balance in Type 2 Diabetes: Role of Purinergic Signalling","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-13 17:49:28","doi":"10.21203/rs.3.rs-7436814/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-11-20T18:01:53+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-20T15:20:58+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-01T14:23:02+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"280605143115738693881162813950226426850","date":"2025-10-24T11:28:37+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"126557955125881320482020186950031489569","date":"2025-10-24T07:18:16+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-18T14:51:26+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"316047386250976708591358310332380093242","date":"2025-09-30T21:42:38+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-30T20:22:24+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-02T09:06:10+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-02T09:06:00+00:00","index":"","fulltext":""},{"type":"submitted","content":"Purinergic Signalling","date":"2025-08-22T18:13:28+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"purinergic-signalling","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pusi","sideBox":"Learn more about [Purinergic Signalling](http://link.springer.com/journal/11302)","snPcode":"11302","submissionUrl":"https://submission.nature.com/new-submission/11302/3","title":"Purinergic Signalling","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"56766987-a367-4096-83d7-6b9157ca2eb2","owner":[],"postedDate":"October 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-03-23T16:07:14+00:00","versionOfRecord":{"articleIdentity":"rs-7436814","link":"https://doi.org/10.1007/s11302-026-10136-8","journal":{"identity":"purinergic-signalling","isVorOnly":false,"title":"Purinergic Signalling"},"publishedOn":"2026-03-17 15:59:41","publishedOnDateReadable":"March 17th, 2026"},"versionCreatedAt":"2025-10-13 17:49:28","video":"","vorDoi":"10.1007/s11302-026-10136-8","vorDoiUrl":"https://doi.org/10.1007/s11302-026-10136-8","workflowStages":[]},"version":"v1","identity":"rs-7436814","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7436814","identity":"rs-7436814","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}