Previous evidence suggests that though N-acetylcysteine (NAC) supplementation following eccentric exercise-induced muscle damage disrupts the skeletal muscle's repair and remodelling process at 8 days of recovery, it attenuates substantially the decline of skeletal muscle performance during the first 48 hours of recovery. The enhanced performance capacity during the first phase of recovery in response to NAC supplementation might be attributed to the altered redox status in skeletal muscle as a consequence of the NAC-mediated elevation of reduced glutathione (GSH) levels. The rise in GSH results in a redox-dependent attenuation of immune cell mobilisation and reduction of oxidative stress response, leading to a blunted rise of muscle damage and inflammatory markers during the first 2-3 days of recovery. However, following exercise-induced muscle damage, protein synthesis increases in skeletal muscle over the next 24-48 hours to support its repair process, and thus protein supplementation might accelerate the recovery process by enhancing the protein synthetic response. Therefore, the present study aims at investigating for first time the combined effect of NAC and whey protein supplementation on the short-term (during the first 72 hours) recovery process of skeletal muscle following damaging exercise (eccentric exercise) and compared it with the well-documented efficacy of NAC supplementation. The results of this study might be particularly useful for athletes, such as those in soccer and basketball, that participate in 3 games or intense training sessions during the same macrocycle with only 48-72 hours of recovery in-between.
Participants will initially undergo baseline testing including assessment of their (i) anthropometrics, (ii) body composition via DXA, (iii) habitual physical activity level via accelerometry, (iv) resting metabolic rate via indirect calorimetry, (v) daily dietary intake via 7-day dietary recalls and (vi) cardiorespiratory fitness using a treadmill ramp test. After baseline testing, participants will be assigned to (a) a Placebo, (b) a N-acetylcysteine (NAC) or (c) a N-acetylcysteine + Whey protein (PRoNAC) group, in a randomized, double-blind, placebo-controlled, parallel group, repeated measures design. In all groups, participants will execute an eccentric exercise protocol on an isokinetic dynamometer consisted of 150 eccentric muscle contractions. Before exercise as well as at 1-, 2-, 3-, 24-, 48- and 72-hours post-exercise they will undergo assessment of their maximal voluntary isometric peal torque, maximal countermovement jump height and muscle soreness level. In addition, blood sample will be drawn before exercise and at 24-, 48- and 72-hours post-exercise for the determination of myoglobin concentration, reduced (GSH) and oxidized (GSSG) glutathione content, protein carbonyl formation and catalase activity. Throughout the 4-day experimental period (Day 1: exercise day, Days 2, 3 and 4: 24-, 48- and 72-hours post-exercise, respectively) participants will receive daily, 3 doses from the respective supplement as follows: the 1st dose at mid-morning (between breakfast and lunch), the 2nd dose at mid-afternoon (post-exercise on day 1 and between lunch and dinner on days 2-4) and the 3rd dose before sleep. In Placebo group, participants will be supplemented with 0.31 gr of maltodextrin per kg of body weight in the first two servings (at mid-morning and mid-afternoon supplements) and with 0.49 gr of maltodextrin per kg of body weight in the pre-sleep serving (1.1 gr maltodextrin/kg BW/day). In NAC group, participants will be supplemented with 13.33 mg of N-acetylcysteine per kg of body weight per serving (40 mg NAC/kg BW/day). In PRoNAC group participants will be supplemented with 0.31 gr of whey protein and 13.33 mg of N-acetylcysteine per kg of body weight in the first two servings (at mid-morning and mid-afternoon supplements) and with 0.49 gr of whey protein and 13.33 mg of N-acetylcysteine per kg of body weight in the pre-sleep serving (1.1 gr Whey protein + 40 mg NAC /kg BW/day).
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
TREATMENT
Masking
TRIPLE
Enrollment
30
Participants will receive three servings of carbohydrate per day during the 4-day experimental period. In specific, they will be supplemented with 0.31 gr of maltodextrin per kg of body weight in the first two servings (at mid-morning and mid-afternoon supplements) and with 0.49 gr of maltodextrin per kg of body weight in the pre-sleep serving (1.1 gr maltodextrin/kg BW/day).
Participants will receive three servings of N-acetylcysteine per day during the 4-day experimental period. In specific, they will be supplemented with 13.33 mg of N-acetylcysteine per kg of body weight per serving (40 mg NAC/kg BW/day).
Participants will receive three servings of N-acetylcysteine + whey protein per day during the 4-day experimental period. In specific, they will be supplemented with 0.31 gr of whey protein and 13.33 mg of N-acetylcysteine per kg of body weight in the first two servings (at mid-morning and mid-afternoon supplements) and with 0.49 gr of whey protein and 13.33 mg of N-acetylcysteine per kg of body weight in the pre-sleep serving (1.1 gr Whey protein + 40 mg NAC /kg BW/day).
Department of Physical Education and Sport Science, University of Thessaly
Trikala, Karies, Greece
RECRUITINGChange in maximal voluntary isometric muscle contraction.
Maximal voluntary isometric muscle contraction will be assessed on an isokinetic dynamometer.
Time frame: Before exercise and at 1-, 2-, 3-, 24-, 48- and 72-hours post-exercise.
Change in countermovement jump height.
Countermovement jump height will be assessed by using the Optojump Next system.
Time frame: Before exercise and at 1-, 2-, 3-, 24-, 48- and 72-hours post-exercise.
Change in delayed onset of muscle soreness (DOMS)
DOMS of the knee extensors of the dominant limb will be evaluated by palpation of the relaxed muscle's belly and distal region with participants rating the perceived soreness on a visual analogue scale ranging from 1 to 10.
Time frame: Before exercise and at 1-, 2-, 3-, 24-, 48- and 72-hours post-exercise.
Change in reduced glutathione content.
Reduced glutathione content in blood erythrocytes will be assessed using spectrophometric method.
Time frame: Before exercise and at 24-, 48- and 72-hours post-exercise.
Change in oxidized glutathione content.
Oxidized glutathione content in blood erythrocytes will be assessed using spectrophometric method.
Time frame: Before exercise and at 24-, 48- and 72-hours post-exercise.
Change in catalase activity.
Catalase activity will be measured spectrophotometrically in red blood cells.
Time frame: Before exercise and at 24-, 48- and 72-hours post-exercise.
Change in protein carbonyl concentration
Protein carbonyl concentration will be measured spectrophotometrically in red blood cells.
Time frame: Before exercise and at 24-, 48- and 72-hours post-exercise.
Change in myoglobin concentration in blood.
Myoglobin concentration in blood will be assessed using an automatic biochemistry analyzer and commercially available kits.
Time frame: Before exercise and at 24-, 48- and 72-hours post-exercise.
Participants' lean body mass
Lean body mass (kg) will be assessed using dual-energy X-ray absorptiometry (DXA) instrumentation.
Time frame: At baseline.
Participants' daily physical activity-related energy expenditure.
Physical activity-related energy expenditure (kcal/day) will be assessed using accelerometers.
Time frame: At baseline.
Participants daily dietary intake.
Daily dietary intake composition (total kcal/day, gr of protein, gr of CHO and gr of fat intake/day, gr/mg of micronutrient intake/day) will be monitored using 24-h dietary recalls that will be analyzed by a trained dietitian using a dietary analysis software.
Time frame: At baseline.
Participants' resting metabolic rate.
Resting metabolic rate will be assessed using indirect calorimetry.
Time frame: At baseline.
Participants' cardiorespiratory fitness status.
Cardiorespiratory fitness will be assessed using a treadmill ramp test.
Time frame: At baseline.
Participants' fat free mass.
Fat free mass will be assessed using dual-energy X-ray absorptiometry (DXA) instrumentation.
Time frame: At baseline.
Participants' fat mass.
Fat mass will be assessed using dual-energy X-ray absorptiometry (DXA) instrumentation.
Time frame: At baseline.
Participants' body fat percent.
Body fat percent (%) will be assessed using dual-energy X-ray absorptiometry (DXA) instrumentation.
Time frame: At baseline.
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