The Effect of Butanediol Ingestion on Skeletal Muscle Angiogenesis in Hypoxia

The Acute Effect of (R)-1,3-butanediol Ingestion on Post-exercise Skeletal Muscle Angiogenesis in Healthy Adults Under Normoxic and Hypoxic Conditions.

This study specifically aims to elucidate the effects of intermittent exogenous ketosis (IEK) as well as hypoxia on muscular pro-angiogenic factors- after a 60-min HIIT bout in normoxia. Moreover, blood and muscle oxygenation status, as well as peripheral blood flow and cognitive function will be assessed.

Study Overview

• Status: Completed
• Conditions: Hypoxia; Angiogenesis; Ketosis
• Intervention / Treatment: Dietary supplement: Normoxic recovery and placebo (NPL); Dietary supplement: Normoxic recovery and ketones (NKE); Dietary supplement: Hypoxic recovery and placebo (HPL); Dietary supplement: Hypoxic recovery and ketones (HKE)

Detailed Description

After an exercise bout, angiogenesis is incredibly important in the recovery process as an increased number of capillaries enables higher metabolite transport to and from the working muscle. Often, a training strategy consists of living-high (sleeping at -stimulated- altitude) and training low (training at sea level). A decreased oxygen availability, also known as hypoxia, however poses an additional stress on the human body, potentially compromising the overall training efficiency. Ketones are recently found to increase angiogenesis in response to overload training (increased skeletal muscle capillarization and VEGF content) and to increase serum EPO concentrations. Therefore, the investigators want to evaluate the isolated and interactive effects of both ketones and hypoxia on post-exercise recovery and factors implicated in skeletal muscle angiogenesis, after training in normoxia. Moreover, a performance test will be performed after 7h of hypoxic or normoxic recovery by means of a simulated 15 min all-out time trial. During the 7h post-exercise window, biological samples are collected (muscle biopsies, venous blood samples, urine, capillary blood samples). Moreover, blood and muscle oxygenation, peripheral blood flow and cognitive function are assessed at regular timepoints.

• Study Type: Interventional
• Enrollment (Actual): 15
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Belgium
  • Vlaams-Brabant
    • Leuven, Vlaams-Brabant, Belgium, 3000
      • KU Leuven

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

1. Written informed consent must be obtained prior to any experimental procedures
2. Biological male or biological females using oral contraception between 18 and 35 years old
3. Recreational or competitive cyclists performing regularly cycling training sessions with an average training volume of more than 6 hours per week
4. Good health status confirmed by a medical screening
5. Body Mass Index (BMI) between 18 and 25 kg/m2

Exclusion Criteria:

1. Any kind of injury/pathology that is a contra-indication for hypoxic exposure and/or to perform high-intensity exercise, evaluated by a sport medical screening
2. Intake of any medication or nutritional supplement that is known to affect exercise or performance. Intake will be assessed during recruitment and the sport medical screening.
3. Intake of analgesics, anti-inflammatory agents, or supplementary anti-oxidants, within two weeks of study participation.
4. Recent residence or training under hypoxia; more than 7 days exposure to altitude > 1500 m during the 3 months preceding the study.
5. Blood donation within 3 months of study participation.
6. Habitual smoking
7. Pre-existing, diagnosed psychiatric conditions or anxiety
8. Females that are pregnant or are planning to be pregnant before the end of the study (end of May 2024)
9. Depression or anxiety as assessed by the Beck Depression Inventory 25 (Appendix 2) and Beck Anxiety Inventory 26 (Appendix 3). Only a score in the range of 'normal ups and downs' (score 1-10) for depression or 'minimal anxiety' (score 0-7) for anxiety are tolerated.
10. History of addiction or excessive caffeine/alcohol consumption assessed by a questionnaire (Appendix 4).
11. Any other reason that might pose undue risk to the participant, or introduce bias into the study outcomes, at the discretion of the research team.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Randomized
• Interventional Model: Crossover Assignment
• Masking: Double

Number of Arms

4

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Placebo Comparator: 1 session with normoxic post-exercise recovery, supplemented with placebo (NPL); Normoxic training, followed by normoxic recovery with placebo supplements	Dietary supplement: Normoxic recovery and placebo (NPL); Altitude: sea level Dietary supplement: placebo. Water: 98.3% w/w Bitter flavour type: 0.7% w/w Monk fruit extract: 0.04% w/w Tasteva: 0.03% w/w Citric acid: 0.6% w/w Flavor blend: 0.02% w/w Bitter masking flavour: 0.2% w/w Passion fruit flavour: 0.005% w/w
Experimental: 1 session with normoxic post-exercise recovery, supplemented with ketones (NKE); Normoxic training, followed by normoxic recovery with ketone supplements	Dietary supplement: Normoxic recovery and ketones (NKE); Altitude: sea level Dietary supplement: Commercially available Ketone-IQ butanediol drink R-1,3-butanediol: 28.6% w/v in water
Experimental: 1 session with hypoxic post-exercise recovery, supplemented with placebo (HPL); Normoxic training, followed by hypoxic (simulated altitude of 3000m) recovery with placebo supplements	Dietary supplement: Hypoxic recovery and placebo (HPL); Altitude: 3,000m (simulated) Dietary supplement: placebo. Water: 98.3% w/w Bitter flavour type: 0.7% w/w Monk fruit extract: 0.04% w/w Tasteva: 0.03% w/w Citric acid: 0.6% w/w Flavor blend: 0.02% w/w Bitter masking flavour: 0.2% w/w Passion fruit flavour: 0.005% w/w
Experimental: 1 session with hypoxic post-exercise recovery, supplemented with ketones (HKE); Normoxic training, followed by hypoxic (simulated altitude of 3000m) recovery with ketone supplements	Dietary supplement: Hypoxic recovery and ketones (HKE); Altitude: 3,000m (simulated) Dietary supplement: Commercially available Ketone-IQ butanediol drink R-1,3-butanediol: 28.6% w/v in water

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Post-exercise muscle VEGF mRNA expression	Measured using PCR on collected muscle biopsies	Muscle biopsies are collected 10 minutes after the end of training as well as 3 hours later.
Post-exercise serum EPO concentrations	Measured using ELISA on collected serum samples	Venous blood samples are collected 10 minutes after the end of training as well as 3 hours, 5 hours and 7 hours later.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Change in muscular VEGF concentration	Measured using Western Blotting on muscle biopsies	Muscle biopsies are collected 10 minutes after the end of training as well as 3 hours later.
Change in serum VEGF concentration	Measured using ELISA on collected serum samples	Venous blood samples are collected immediately (5 minutes) after the end of training, as well as 3hours, 5hours and 7hours later..
Change in blood oxygenation	Measured using pulse oximetry with a sensor on the forehead	Continuously measured starting 30 minutes after the end of exerciseuntil 7.5 hours later
Exercise performance	Measured as the average power output (W) during a 15 minutes all-out time trial	Performed 7hours and 30minutes after the end of training
Change in skeletal muscle oxygenation	Measured using NIRS with a sensor on the belly of the m. vastus lateralis (skeletal muscle)	Measured during resting measurements performed 30 minutes, 3hours and 30minutes as well as 6hours and 30minutes after the end of training.
Change in peripheral blood flow	Measured using duplex ultrasonography	Measured during resting measurements performed 30 minutes, 3hours and 30minutes as well as 6hours and 30minutes after the end of the training

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Change in citrate synthase as a marker of mitochondrial density	Measured using an enzymatic essay	Muscle biopsies are collected 10 minutes after the end of training as well as 3 hours later.

Collaborators and Investigators

• Sponsor: KU Leuven
• Investigators: Principal Investigator: Chiel Poffé, KU Leuven

Study record dates

Study Major Dates

• Study Start (Actual): July 1, 2024
• Primary Completion (Actual): July 30, 2024
• Study Completion (Actual): July 30, 2024

Study Registration Dates

• First Submitted: May 30, 2024
• First Submitted That Met QC Criteria: June 18, 2024
• First Posted (Actual): June 25, 2024

Study Record Updates

• Last Update Posted (Actual): September 19, 2024
• Last Update Submitted That Met QC Criteria: September 12, 2024
• Last Verified: September 1, 2024

More Information

Terms related to this study

• Keywords: Exercise; Recovery; Living high training low
• Additional Relevant MeSH Terms: Signs and Symptoms, Respiratory; Hypoxia
• Other Study ID Numbers: S68402

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No