Redox Regulation of Satellite Cells and Skeletal Muscle Healing

Redox Status Clinical Trial

Official title:

The Effect of Redox Potential on the Regulation of Satellite Cells and Skeletal Muscle Healing Following Exercise-Induced Muscle Damage

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT03711838
• Other study ID #: Satellite Cells - UTH
• Status: Completed
• Phase: N/A
• Start date: July 22, 2019
• Est. completion date: August 1, 2022

Study information

• Verified date: December 2022
• Source: University of Thessaly
• Contact: n/a
• Is FDA regulated: No
• Study type: Interventional

Clinical Trial Summary

Skeletal muscle stem cells (Satellite cells) are indispensable for muscle growth and remodeling following myofibril damage. Skeletal muscle trauma is present in numerous catabolic conditions, characterized by elevated proteolysis and muscle wasting such as, cancer cachexia and muscular dystrophy, which result in physical capacity impairment and a deteriorated quality of life. Recent studies performed in animals and cell cultures indicate that the increased levels of inflammation and oxidative stress and the reduction of antioxidant defense may blunt the satellite cells response and myogenic programming during muscle healing. However, evidence regarding the effects of redox status on satellite cells and muscle myogenic potential in humans is lacking. Exercise-induced muscle damage bears striking similarities with the aforementioned conditions, which makes it a valuable tool to investigate the redox-dependent regulation of satellite cells during muscle healing. Thus, the objectives of the present study are to examine the effects of redox status perturbation (via N-acetylcysteine administration) on intracellular pathways responsible for satellite cells responses at rest and following aseptic muscle trauma induced by damaging exercise.

Description:

A total number of 40-60 males, young individuals aged 18-30 years, will be initially enrolled in the study. Then, participants will be allocated to either a 1) Low Respondents (LR) or a 2) High Respondents (HR) group based on the basal satellite cells content of their vastus lateralis muscle of their dominant leg. In a double-blind, crossover, repeated measures design, participants will consume either Placebo (PLA) or N-acetylcysteine (NAC) before (7-day loading phase), on exercise day and for 8 consecutive days following a single bout of intense exercise (300 eccentric contractions at 30 deg/sec in an isokinetic dynamometer). In both conditions, blood samples and muscle biopsies will be collected at baseline, before the exercise protocol and at 2- and 8-days post-exercise. Muscle performance and soreness will also be assessed at the same time points. Before the first trial, participants' dietary intake (via diet recalls) and physical activity (via accelerometry) will be analyzed and nutritional plans will be provided to participants in order to normalize their antioxidant and protein intake. A 4-week washout period will be implemented between trials. Blood samples will be analyzed for inflammation and oxidative stress markers. Muscle samples will be analyzed for satellite cells responses and myogenic potential, protein levels of intracellular signaling proteins, muscle thiols and antioxidant enzymes activity.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 45
• Est. completion date: August 1, 2022
• Est. primary completion date: May 1, 2022
• Accepts healthy volunteers: Accepts Healthy Volunteers
• Gender: Male
• Age group: 18 Years to 30 Years

Eligibility

Inclusion Criteria:

1. No recent history of musculoskeletal injury

2. Non-smokers.

3. Abstain from any vigorous physical activity during the study

4. Abstain from consumption of caffeine, alcohol, performance-enhancing or antioxidant supplements, NSAIDs and medications before (at least 6 months) and during the study.

Exclusion Criteria:

1. A known NAC intolerance or allergy

2. A recent febrile illness

3. A recent history of muscle lesion and/or lower limb trauma

4. Presence of metabolic diseases

5. Use of anti-inflammatory medication.

6. Use of medication interacting with muscle metabolism.

Related Conditions & MeSH terms

• Aseptic Inflammation
• Exercise-induced Muscle Trauma
• Inflammation
• Redox Status
• Satellite Cells

Intervention

Dietary Supplement:
• N-Acetylcysteine
N-Acetylcysteine in a powder form diluted in a 250 ml drink containing 248 ml water and 2 ml of natural, non-caloric, flavoring-sweetener containing sucralose.
• Placebo
Placebo consisted of 248 ml water and 2 ml of natural, non-caloric, flavoring-sweetener containing sucralose.

Locations

Country	Name	City	State
Greece	Laboratory of Exercise Biochemistry, Exercise Physiology,and Sports Nutrition, School of Physical Education and Sport Science, University of Thessaly	Trikala	Thessaly

Sponsors (1)

Lead Sponsor	Collaborator
University of Thessaly

Country where clinical trial is conducted

Greece, 

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Change in muscle satellite cells number (i.e. Pax7+ cells) and activation status (i.e. Pax7+/MyoD+ cells)	Satellite cells number and activation status, will be assessed in muscle via immunohistochemistry.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Primary	Change in muscle myogenic mRNA expression	mRNA expression levels of Myogenic factor 5 (Myf5), myogenin and Myogenic factor 6 (Myf6/MRF4) and myostatin will be assessed in muscle using Real-Time Polymerase Chain Reaction (RT-PCR).	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Primary	Change in muscle inflammatory state	Pro-inflammatory (M1+) and anti-inflammatory (M2+) macrophages will be measured in muscle using immunohistochemistry.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Primary	Change in intracellular antioxidant enzymes in muscle	Protein levels of Glutathione peroxidase 3 (GPx3), Superoxide dismutase 1 (SOD1) and Thioredoxin (Trx1) will be measured using western blotting.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Primary	Change in muscle thiol content	Concentration levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) will be measured spectophotometrically.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Primary	Change in intracellular signaling proteins in muscle	Protein levels of IGF-1, Notch1 and Wnt3 will be measured using western blotting.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Primary	Change in skeletal muscle damage levels	Skeletal muscle damage will be quantified via histochemistry using H&E staining.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Resting metabolic rate (RMR)	RMR will be assessed after an overnight fast with participants in a supine position following a 15-min stabilization period by taking 30 consecutive 1-min VO2/CO2 measurements using a portable open-circuit indirect calorimeter with a ventilated hood system following a standard calibration protocol.	At baseline
Secondary	Body composition	Body composition will be measured using a dual-energy x-ray absorptiometry scanner (DXA).	At baseline
Secondary	Maximal oxygen consumption (VO2max)	VO2max will be assessed during continuous incremental running to volitional fatigue on a treadmill with a pulmonary gas exchange system (Oxycon Mobile; Sensor-Medics Corporation).	At baseline
Secondary	Isokinetic strength	Maximal knee extensor eccentric peak torque at 60 degrees will be assessed on an isokinetic dynamometer.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Change in delayed onset of muscle soreness (DOMS)	Muscle soreness will assessed during palpation of the muscle belly and the distal region of relaxed vastus medialis, vastus lateralis and rectus femoris following three repetitions of of a full squat. Subjects will rate their DOMS on a visual analogue scale (0-10).	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Physical activity	Level of habitual physical activity will be assessed using accelerometry (ActiGraph GT3X-BT accelerometer).	At baseline.
Secondary	Dietary intake	Daily dietary intake will be assessed using 7-day diet recalls.	At baseline.
Secondary	Interleukin-1ß (IL-1ß) in blood	Concentration of IL-1ß will be measured in plasma.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Interleukin-6 (IL-6) in blood	Concentration of IL-6 will be measured in plasma.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Interleukin-8 (IL-8) in blood	Concentration of IL-6 will be measured in plasma.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Interleukin-10 (IL-10) in blood	Concentration of IL-10 will be measured in plasma.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Cortisol in blood	Concentration of cortisol will be measured in serum.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Protein carbonyls (PC)	Concentration of protein carbonyls will be measured in muscle.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Malondialdehyde (MDA)	Concentration of malondialdehyde will be measured in muscle.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Total antioxidant capacity (TAC)	Concentration of TAC will be measured in serum.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Catalase	Concentration of catalase will be measured in red blood cell lysate.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	Creatine Kinase (CK)	Concentration of CK will be measured in serum.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.
Secondary	White blood cell count in blood	White blood cell count will be measured in blood. White blood cell count will be measured in blood.	At baseline, before the exercise protocol and at days 2 and 8 following exercise.