Antioxidants Combined With Cryotherapy on Inflammatory Response After Resistance Exercise in Untrained Volunteers

Study Overview
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Brief Summary
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Acutely resistance exercise induces inflammatory responses and leukocytosis arising of oxidative stress, that clinically manifested by pain and/or delayed onset muscle soreness (DOMS). Beginners in resistance exercise programs are more vulnerable to the effects of oxidative stress as they exhibit lower antioxidant capacity, greater lipid peroxidation and present increased perception of pain after exercises that may lead to abandonment the exercises practice. Vitamins C and E are exogenous antioxidants which are able to prevent damages caused by oxidative stress. Cryotherapy decreases temperature and reduced generation of reactive oxygen species. The aim of the present research are to investigate the effects of the concomitant of vitamins and of cryotherapy on leukocytosis, inflammatory markers, oxidative stress parameters and pain in untrained individuals submitted to a resistance exercise session.

Detailed Description
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The regular practice of physical exercises promotes anti-inflammatory effects, which reduces the mortality from all causes, especially by cardiovascular diseases. On the other hand, physical exercise induces acute inflammatory responses. Acute resistance exercises provoke mechanical and metabolic stress which may vary depending on the intensity, specificity, volume and workload. During these exercises, reactive oxygen and nitrogen species (RONS) are formed and, when its production exceeds the antioxidant enzymatic capacity (Superoxide dismutase, Catalase and Glutathione peroxidase) and non-enzymatic capacity (vitamins A, C, E and uric acid) results in oxidation of cell constituents. This state of oxidative stress promotes one inflammatory response, activation and mobilization of circulating white blood cells and induce a transitory leukocytosis, that are observed during and immediately after resistance exercises. The local neutrophilia induces the amplification of inflammatory response by feedback resulting in activation and/or mobilization of the more white blood cells. This sequence of physiological events leads to muscular remodeling although, during such process, occur functional insufficiency and the sensation of pain and/or delayed onset muscular soreness (DOMS), which deserves special attention, once may lead to interruption or abandonment of exercises, especially concerning beginners in this practice. Numerous preventive approaches have been studied intending to minimize the inflammatory response caused by resistance exercises, including the supplementation with vitamins C (ascorbic acid) and E (alfa-tocopherol). These vitamins are exogenous antioxidants which are able to prevent damages caused by RONS and consequently attenuate the oxidative damage induced by resistance exercises. Cryotherapy is defined as the therapeutic application of any substance that removes heat from the body, thus lowering the temperature of tissues. Immersion in cold water (≤ 15ºC) is a common method of heat reduction. Cryotherapy attenuates microvascular dysfunction, and decreases temperature, metabolism and oxygen demand in the electron transport chain. As a consequence, lower amounts of RONS are produced less damage was caused to adjacent molecules. The aim of the present research are to investigate the effects of the concomitant supplementation of vitamins (C and E) and cryotherapy (immersion in water at 15ºC) on leukocytosis, inflammatory markers, oxidative stress parameters and pain in untrained individuals submitted to a resistance exercise session.

Official Title
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Vitamins C and E Combined With Cryotherapy on the Pain, Oxidative Stress and Inflammatory Response After Resistance Exercise Session in Untrained Volunteers

Conditions
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Pain, Inflammation, Oxidative Stress

Intervention / Treatment
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* Other: Cryotherapy
* Other: Vitamins C and E
* Other: Vitamins C and E associated with cryotherapy
* Other: Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Age between 20 and 35 years, body mass index (MBI: kg/m2) lower than 30, non-smokers, did not practice physical and/or regular physical exercises (less than two times per week), presented no previous diagnosis of chronic diseases (rheumatic, cardiovascular, metabolic, neurological, oncologic, immunological or hematologic disorders), is not engaged in any diet programs and is not making use of medication. Exclusion Criteria: Inflammatory response (ultrasensitive C reactive protein >3 mg/dL), hyperglycemia (>100 mg/dL), leukocytosis, hyperthermia (>38ºC), changes in the systemic blood pressure (>140/90 mmHg) and/or any symptom of pain or discomfort

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 35 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Prevention
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: 1ª Session of resistance exercises<br>Exercise session are based on previous studies (TEIXEIRA et al., 2012; TEIXEIRA et al., 2014) and will be randomized according to the sequence of exercises (extensor bench, squat and leg press) and the interventions by drawing sealed brown envelopes. Forty minutes before basal blood collection and of the exercise session the volunteers will receive placebo (two pills wheat flour) and the remaining procedures will be conserved. The exercise sessions were comprised of four series of 10 maximum repetitions, with an interval of one minute between series and two minutes between exercises. Before the 10 maximum repetitions test and data collection, standard instructions will be given concerning the experimental procedure and execution technique of the exercises. | Other: Cryotherapy<br>* The intervention will consist of the application of hypothermia by immersion of the lower limbs of water on individuals to 15ºC for a period of 10 minutes immediately after the exercise protocol.<br>* Other names: Hypothermia;Other: Vitamins C and E<br>* The session of resistance exercises with intervention will be based on the supplementation with vitamin C (1000 mg) and vitamin E (800 IU) by oral intake (100mL of water) 40 minutes before basal blood collection.<br>* Other names: alfa-tocopherol;Other: Vitamins C and E associated with cryotherapy<br>* The session of resistance exercises with intervention will be based on the supplementation with vitamin C (1000 mg) and vitamin E (800 IU) by oral intake (100mL of water) 40 minutes before basal blood collection. The intervention will consist of the application of hypothermia by immersion of the lower limbs of water on individuals to 15ºC for a period of 10 minutes immediately after the exercise protocol.<br>* Other names: Hypothermia;Other: Placebo<br>* The volunteers from placebo exercise session will receive two pills (containing wheat flour) and the remaining procedures were conserved.<br>* Other names: Control;|
| Active Comparator: 2ª Session of resistance exercises<br>Exercise session are based on previous studies (TEIXEIRA et al., 2012; TEIXEIRA et al., 2014) and will be randomized according to the sequence of exercises (extensor bench, squat and leg press) and the interventions by drawing sealed brown envelopes. Forty minutes before basal blood collection and of the exercise session the volunteers will receive placebo (two pills wheat flour) and the remaining procedures will be conserved. The exercise sessions were comprised of four series of 10 maximum repetitions, with an interval of one minute between series and two minutes between exercises. Before the 10 maximum repetitions test and data collection, standard instructions will be given concerning the experimental procedure and execution technique of the exercises. | Other: Cryotherapy<br>* The intervention will consist of the application of hypothermia by immersion of the lower limbs of water on individuals to 15ºC for a period of 10 minutes immediately after the exercise protocol.<br>* Other names: Hypothermia;Other: Vitamins C and E<br>* The session of resistance exercises with intervention will be based on the supplementation with vitamin C (1000 mg) and vitamin E (800 IU) by oral intake (100mL of water) 40 minutes before basal blood collection.<br>* Other names: alfa-tocopherol;Other: Vitamins C and E associated with cryotherapy<br>* The session of resistance exercises with intervention will be based on the supplementation with vitamin C (1000 mg) and vitamin E (800 IU) by oral intake (100mL of water) 40 minutes before basal blood collection. The intervention will consist of the application of hypothermia by immersion of the lower limbs of water on individuals to 15ºC for a period of 10 minutes immediately after the exercise protocol.<br>* Other names: Hypothermia;Other: Placebo<br>* The volunteers from placebo exercise session will receive two pills (containing wheat flour) and the remaining procedures were conserved.<br>* Other names: Control;|
| Active Comparator: 3ª Session of resistance exercises<br>Exercise session are based on previous studies (TEIXEIRA et al., 2012; TEIXEIRA et al., 2014) and will be randomized according to the sequence of exercises (extensor bench, squat and leg press) and the interventions by drawing sealed brown envelopes. Forty minutes before basal blood collection and of the exercise session the volunteers will receive placebo (two pills wheat flour) and the remaining procedures will be conserved. The exercise sessions were comprised of four series of 10 maximum repetitions, with an interval of one minute between series and two minutes between exercises. Before the 10 maximum repetitions test and data collection, standard instructions will be given concerning the experimental procedure and execution technique of the exercises. | Other: Cryotherapy<br>* The intervention will consist of the application of hypothermia by immersion of the lower limbs of water on individuals to 15ºC for a period of 10 minutes immediately after the exercise protocol.<br>* Other names: Hypothermia;Other: Vitamins C and E<br>* The session of resistance exercises with intervention will be based on the supplementation with vitamin C (1000 mg) and vitamin E (800 IU) by oral intake (100mL of water) 40 minutes before basal blood collection.<br>* Other names: alfa-tocopherol;Other: Vitamins C and E associated with cryotherapy<br>* The session of resistance exercises with intervention will be based on the supplementation with vitamin C (1000 mg) and vitamin E (800 IU) by oral intake (100mL of water) 40 minutes before basal blood collection. The intervention will consist of the application of hypothermia by immersion of the lower limbs of water on individuals to 15ºC for a period of 10 minutes immediately after the exercise protocol.<br>* Other names: Hypothermia;Other: Placebo<br>* The volunteers from placebo exercise session will receive two pills (containing wheat flour) and the remaining procedures were conserved.<br>* Other names: Control;|
| Active Comparator: 4ª Session of resistance exercises<br>Exercise session are based on previous studies (TEIXEIRA et al., 2012; TEIXEIRA et al., 2014) and will be randomized according to the sequence of exercises (extensor bench, squat and leg press) and the interventions by drawing sealed brown envelopes. Forty minutes before basal blood collection and of the exercise session the volunteers will receive placebo (two pills wheat flour) and the remaining procedures will be conserved. The exercise sessions were comprised of four series of 10 maximum repetitions, with an interval of one minute between series and two minutes between exercises. Before the 10 maximum repetitions test and data collection, standard instructions will be given concerning the experimental procedure and execution technique of the exercises. | Other: Cryotherapy<br>* The intervention will consist of the application of hypothermia by immersion of the lower limbs of water on individuals to 15ºC for a period of 10 minutes immediately after the exercise protocol.<br>* Other names: Hypothermia;Other: Vitamins C and E<br>* The session of resistance exercises with intervention will be based on the supplementation with vitamin C (1000 mg) and vitamin E (800 IU) by oral intake (100mL of water) 40 minutes before basal blood collection.<br>* Other names: alfa-tocopherol;Other: Vitamins C and E associated with cryotherapy<br>* The session of resistance exercises with intervention will be based on the supplementation with vitamin C (1000 mg) and vitamin E (800 IU) by oral intake (100mL of water) 40 minutes before basal blood collection. The intervention will consist of the application of hypothermia by immersion of the lower limbs of water on individuals to 15ºC for a period of 10 minutes immediately after the exercise protocol.<br>* Other names: Hypothermia;Other: Placebo<br>* The volunteers from placebo exercise session will receive two pills (containing wheat flour) and the remaining procedures were conserved.<br>* Other names: Control;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Delayed onset muscle soreness (points) | Twenty-four hours after the exercise session, the pain or delayed onset muscle soreness (DOMS) was evaluated by visual analog scale (0 - 10 points). | Assessed 24 hours after resistance exercise session | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Erythrocytes (/ mm3) | Blood count (/ mm3). | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 
| Platelets (/ mm3) | Blood count (/ mm3). | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 
| Leukogram (/ mm3) | Differential white blood cell counts (/ mm3). Total leukocytes, segmented neutrophils, young neutrophils (rods), monocytes, lymphocytes and eosinophils. | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 
| C reactive protein (mg/dL) | Inflammatory markers | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 
| Creatine kinase (U/L) | Inflammatory markers | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 
| Lactate dehydrogenase (mmol/L) | Inflammatory markers | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 
| Fibrinogen (mg/dL) | Inflammatory markers. | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 
| Reactive oxygen species (ROS = relative area) | Oxidative stress parameters: damage molecular. | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 
| Anti-oxidant capacity against peroxyl radicals (ACAP = deference of the areas (wtih ABAP / without ABAP) | Oxidative stress parameters: damage molecular , anti-oxidant capacity against peroxyl radicals (ACAP = deference of the areas (wtih ABAP / without ABAP) | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 
| Superoxide dismutase (SOD = units / mg protein) | Oxidative stress parameters: enzymatic molecular defense | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 
| Catalase (units / mg protein) | Oxidative stress parameters: enzymatic molecular defense | Baseline (before), and at 0, 30 and 120 minutes after exercises. | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Muscle soreness, Free radicals, Ascorbic acid, Alpha-Tocopherol, Cryotherapy, Exercise tolerance, Resistance training