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Completed

NCT Number: NCT06054607

Effect of Short-chain Fatty Acids on Aerobic Endurance

Randomized, double-blind, placebo-controlled crossover study designed to determine the effects of increasing colonic short-chain fatty acid (SCFA) content on aerobic endurance in healthy adults, and to identify underpinning mechanisms. In random order, healthy physically active adults will consume provided diets low in fiber and supplemented with SCFA-enriched high amylose maize starch (a poorly digested resistant starch considered a fermentable fiber) or low amylose maize starch (a rapidly digestible starch) for 1-week separated by a ≥2-week washout. At the end of each intervention period, participants will complete an endurance exercise bout followed by a time trial. Biological samples will be collected to assess muscle and whole body metabolism, gut microbiota, inflammation, and gastrointestinal function.

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Key information

Age range

17 year–39 year

Sex eligibility

All sexes

Study type

Interventional

Phase

Not applicable

Primary location

United States Army Research Institute of Environmental Medicine

Natick, Massachusetts, 01760, United States

Who can participate

Healthy volunteers accepted: Yes

Only the study team can determine whether someone qualifies for participation.

Inclusion criteria

  • Men and women aged 18 - 39 years (active-duty personnel who are 17 yr of age will also be allowed to participate).
  • In good health.
  • Routinely participate in moderate or higher intensity aerobic and/or resistance exercise at least 4 days per week for ≥20 min/d.
  • Meet Army weight for height and body composition standards as defined in Army Regulation 600-9.
  • Self-reports a usual bowel movement frequency of every other day or more often.
  • Willing to refrain from the use of caffeine, alcohol, and nicotine while consuming study diets.
  • Willing to refrain from all dietary supplements beginning 2 weeks prior to study start and throughout study participation.
  • Willing to refrain from consumption of any foods containing live microorganisms (e.g., yogurt, kefir, kombucha) or added prebiotics (e.g. Fiber One products) beginning 2 weeks prior to study start and throughout study participation.
  • Willing to participate in all study procedures.
  • Females must have normal menstrual cycles between 26-32 days in duration; 5 menstrual cycles within the past 6 months; or be using an oral/hormonal contraceptive which contains low-dose estrogen/progesterone to maintain continuous levels throughout the 28-day cycle (i.e., no placebos).

Exclusion criteria

  • Females who are pregnant, expecting to become pregnant during the study, or breastfeeding.
  • Any of the following medical conditions:

Cardiac disease (including arrhythmia or fast or skipped heart beats) Hypertension Musculoskeletal injuries that compromise exercise capability Metabolic or cardiovascular abnormalities (e.g., kidney disease, diabetes, etc.) Disease of the GI tract including, but not limited to diverticulitis, inflammatory bowel disease, irritable bowel syndrome, peptic ulcer disease, Crohn's disease, and ulcerative colitis Allergy to skin adhesive or Lidocaine (or other local anesthetic being used in place of Lidocaine) Suspected or known strictures, fistulas, or physiological/mechanical GI obstruction History of gastric bezoar Swallowing disorders; severe dysphagia to food or pills Implanted or portable electro-mechanical medical devices (e.g., pacemaker)

  • Colonoscopy within 3 months of study participation.
  • Any use of antibiotics or antimycotics, except topical antibiotics/antimycotics, within 3 months of study participation.
  • Regular (i.e., weekly or more frequent) use of over-the-counter medications (including antacids, laxatives, stool softeners, and anti-diarrheals) unless approved by study PI.
  • Use of medication (i.e., diabetes medications, statins, corticosteroids, etc.) that affects macronutrient utilization and/or the ability to participate in strenuous exercise.
  • Anemia (HCT <38 for men and <34 for women) or Sickle Cell Anemia/Trait.
  • Not willing or able to follow all study procedures and diet/exercise restrictions.
  • Allergies, intolerances, unwillingness or inability to eat provided foods and beverages.
  • Following vegetarian/vegan diet or other highly restrictive diet (e.g., ketogenic diet, very high protein diet, Paleo diet, etc.).
  • Any previous blood donation within the previous 8 weeks that when combined with the volume of blood collected for the study within any 8-week period would exceed 550mL.

Treatment and study plan

High-amylose maize starch+acetate/butyrate

Dietary Supplement

High-amylose maize starch (HAMS), commercially available as Hylon VII (Ingredion, Inc.), to which the SCFA acetate or butyrate has been chemically added. Hylon VII is a HAMS containing ~70% amylose and used in a variety of food products. To produce SCFA-enriched HAMS, Hylon VII is chemically modified through esterification with the SCFA acetate or butyrate.

Low-amylose maize starch

Other

Rapidly digestible low-amylose maize starch (0% amylose, 100% amylopectin) sold commercially as AMIOCA (Ingredion, Inc).

Primary outcomes

  1. Run time

    Time frame: Phases 1 and 2, day 8

    Time to complete a 5km treadmill run

Secondary outcomes

  1. Muscle glycogen content

    Time frame: Phases 1 and 2, day 8

    Muscle glycogen content

  2. Plasma glucose turnover

    Time frame: Phases 1 and 2, day 8

    Glucose production, utilization and metabolic clearance rate measured during endurance steady state exercise using 6,6-[2H2] glucose tracer

  3. Whole body substrate oxidation

    Time frame: Phase 1 and 2, day 8

    Carbohydrate and fat oxidation rates measured during endurance steady state exercise using indirect calorimetry

  4. Serum short-chain fatty acid concentrations

    Time frame: Phases 1 and 2, day 8. Measured fasted (-300 minutes) and before (0 minutes) and at the end (80 minutes) of steady state endurance exercise

    Serum acetate, butyrate and propionate concentrations

  5. Fecal short-chain fatty acid concentrations

    Time frame: Phases 1 and 2, days 1 and 7

    Fecal acetate, butyrate and propionate concentrations

  6. Intestinal permeability

    Time frame: Phases 1 and 2, day 8

    Small and large intestine permeability measured by saccharide excretion in urine

Other outcomes

  1. Gastrointestinal symptoms

    Time frame: Phases 1 and 2, daily on days 1 through 8

    Gastrointestinal symptoms (bloating, pain, nausea, flatulence, discomfort, urge to defecate) measured by 10 cm (0 = low, 10 = high) visual analog scale

  2. Circulating biomarkers of substrate utilization

    Time frame: Phases 1 and 2, day 8. Measured before (-300minutes, 0minutes) and during (20, 40, 60, 80minutes) steady state endurance exercise.

    Serum/plasma glucose, insulin, free fatty acids, glycerol, lactate and beta-hydroxybutyrate

  3. Total protein content in muscle

    Time frame: Phases 1 and 2, day 8

    Total protein content measured in muscle collected before and after steady state endurance exercise.

  4. Total protein expression of molecular markers associated with regulation of muscle metabolism.

    Time frame: Phases 1 and 2, day 8

    Muscle protein signalling measured by Western blotting in muscle collected before and after steady state endurance exercise. Total protein expression of molecular markers associated with regulation of muscle metabolism will be measured to include (but not be limited to): Akt, p-AktSer473, AMPKα, p- AMPKαThr172, PGC-1α , SIRT1, ACC, p-ACCSer79, p53, p-p53Ser15, PDK4, IRS1, p-IRS1Ser302, GSK-3β, p-GSK-3βSer9, GSK-3α , p-GSK-3α Ser21, GS, p-GSSer641, and GLUT4.

  5. Phosphorylation status of molecular markers associated with regulation of muscle metabolism.

    Time frame: Phases 1 and 2, day 8

    Muscle protein signalling measured by Western blotting in muscle collected before and after steady state endurance exercise. Phosphorylation status of molecular markers associated with regulation of muscle metabolism will be measured to include (but not be limited to): Akt, p-AktSer473, AMPKα, p- AMPKαThr172, PGC-1α , SIRT1, ACC, p-ACCSer79, p53, p-p53Ser15, PDK4, IRS1, p-IRS1Ser302, GSK-3β, p-GSK-3βSer9, GSK-3α , p-GSK-3α Ser21, GS, p-GSSer641, and GLUT4 .

  6. Pyruvate dehydrogenase activity in muscle

    Time frame: Phases 1 and 2, day 8

    Pyruvate dehydrogenase activity in muscle before and after steady state endurance exercise measured colorimetric assay.

  7. Gene expression

    Time frame: Phases 1 and 2, day 8

    Expression of genes regulating carbohydrate and fat oxidation in muscle collected before and after steady state endurance exercise. Individual primers will be used to determine the mRNA expression of known intracellular targets regulating substrate metabolism to include (but not limited to): FAT, PDK4, HADHA, PGC-1α, PPARδ, PPARγ, FABP, CPT1a, ACOX1, GS, GSK3α, GLUT4, and HK2.

  8. microRNA expression

    Time frame: Phases 1 and 2, day 8

    Expression of genes and microRNA regulating carbohydrate and fat oxidation in muscle collected before and after steady state endurance exercise. microRNA analysis will be conducted using microarray, assessing microRNA that may be associated with substrate metabolism.

  9. Gut microbiota composition

    Time frame: Phases 1 and 2, days 1 and 7

    Relative abundance of bacterial taxa measured by 16S rRNA amplicon sequencing

  10. Gut microbiome gene content

    Time frame: Phases 1 and 2, days 1 and 7

    Relative abundances of microbial genes measured by shotgun metagenomics sequencing

  11. Circulating biomarkers of inflammation

    Time frame: Phases 1 and 2, day 8. Measured before (-300minutes) and at the end (80minutes) of steady state endurance exercise.

    Serum interleukin (IL)-6, IL-10, IL-17, IL-8, IL-1ralpha, IL-1beta, tumor necrosis factor-alpha, interferon gamma

  12. Circulating biomarkers of gut barrier function

    Time frame: Phases 1 and 2, day 8. Measured fasted (-300minutes) and before (0minutes), during (40minutes) and at the end (80minutes) of steady state endurance exercise.

    Serum claudin-3, intestinal fatty acid binding protein and zonulin

  13. Circulating microRNA

    Time frame: Phases 1 and 2, day 8. Measured fasted (-15minutes) and at the end (80minutes) of steady state endurance exercise.

    Serum C-microRNA

  14. Appetite

    Time frame: Phases 1 and 2, daily on days 1 through 7

    Appetite (hunger, fullness, desire to eat, prospective consumption) measured by 10 cm (0 = low, 10 = high) visual analog scale

  15. Breath hydrogen and methane

    Time frame: Phases 1 and 2, day 8. Measured fasting (-300minutes) and before (0minutes) and at the end (80minutes) of steady state endurance exercise

    Breath hydrogen and methane concentrations

Sponsors and collaborators

Lead sponsor

United States Army Research Institute of Environmental Medicine

Fed

Registry information

Important dates

Study start
2023
Primary completion
2025
Study completion
2025
First posted
Sep 26, 2023
Registry last updated
Dec 26, 2025

OpenTrials presents study information sourced from ClinicalTrials.gov. The official registry record should be consulted for the latest information.

View the official ClinicalTrials.gov record (opens in a new tab)

This listing is for discovery and informational purposes only. It is not medical advice, does not guarantee that a study is recruiting, and does not determine eligibility. Contact the study team and a qualified healthcare professional when considering participation.

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