Exercise Training Intensity and Nitrate Supplementation in Post-Menopausal Females

Effects of Exercise Training Intensity and Inorganic Nitrate Supplementation on Vascular Health and Fitness in Post-Menopausal Females

Menopause greatly increases cardiovascular disease risk (CVD). Current exercise guidelines are inadequate to ameliorate this risk and higher intensity exercise may be necessary for cardiovascular benefits in postmenopausal females. Oral nitrate supplementation enhances exercise performance and CVD risk profile in several clinical conditions. The investigators recently reported that 3-days of nitrate supplementation in postmenopausal females enhances acute post-exercise vascular function, in an intensity dependent manner. The effects of nitrate supplementation and exercise training over a longer training program remains unexplored. This investigation will examine the impact of exercise training intensity with and without inorganic nitrate supplementation, on CVD and fitness outcomes. Postmenopausal females will be tested for maximal oxygen uptake and lactate threshold before randomization to one of four groups: that inorganic nitrate-rich beetroot juice, or beetroot juice with nitrate extracted; and assigned to one-month of calorie matched high-intensity or moderate-intensity exercise training. Vascular function and fitness will be evaluated before and after training.

Study Overview

• Status: Recruiting
• Conditions: Menopause
• Intervention / Treatment: Behavioral: High Intensity Exercise; Behavioral: Moderate Intensity Exercise; Dietary supplement: Nitrate-Rich Beetroot Juice; Dietary supplement: Nitrate-Depleted Placebo Beetroot Juice

• Study Type: Interventional
• Enrollment (Estimated): 40
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Casey Derella, PhD; Phone Number: 434-924-1655; Email: bxg7vn@uvahealth.org

Study Locations

• United States
  • Virginia
    • Charlottesville, Virginia, United States, 22903
      • Recruiting
      • Student Health and Wellness Building - Department of Kinesiology
      • Contact: Casey Derella, PhD; Phone Number: 434-924-1655; Email: bxg7vn@uvahealth.org

Participation Criteria

Eligibility Criteria

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

Description

Inclusion Criteria:

• Post-menopausal female (greater than age 45 but less than age 75)
• Sedentary (does not exercise regularly or less than 2 bouts of exercise per week)
• No major changes in medication in the last 3 months

Exclusion Criteria:

• Smokers within last 5 years
• Weight unstable (loss/gain of more than 3kg in the past 3 months)
• Any medical condition that prevents the subject from exercising safely
• Hormone replacement therapy (current or within last 3 months)
• Diabetes
• Currently or recently on vasoactive medications (i.e., calcium channel blockers, statins, ACE or renin inhibitors, ARBs, nitrates, alpha- or beta-blockers, diuretics, proton pump inhibitors, etc.)
• Oral antibiotic use within previous four weeks
• Oral disease or poor oral health as determined by the Oral Health Questionnaire
• Using an antibacterial mouthwash or a mouthwash containing chlorhexidine and unwilling to discontinue use

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Supportive Care
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Quadruple

Number of Arms

4

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Nitrate-Rich Beetroot Juice + High Intensity Exercise (HIE); Participants assigned to this group will be instructed to consume 1 bottle of nitrate-rich beetroot juice (~400mg of nitrate each) 2x a day (morning a night) starting 2-hours before the first exercise session. Participants will complete a supervised bout of exercise on a treadmill 3x a week for 1-month. Participants will exercise at an RPE of 15-17 based on the participant's VO2peak test.	Behavioral: High Intensity Exercise; exercise at an intensity midway between the lactate threshold and VO2peak, from the screening test at an RPE of ~ 15-17; Dietary supplement: Nitrate-Rich Beetroot Juice; subjects will consume 1 bottle of nitrate-rich beetroot juice (~400mg of nitrate each) 2x a day; Other Names: Inorganic Nitrate; James White Beet It
Placebo Comparator: Nitrate-Depleted Placebo Beetroot Juice + High Intensity Exercise (HIE); Participants assigned to this group will be instructed to consume 1 bottle of nitrate-depleted beetroot juice 2x a day (morning a night) starting 2-hours before the first exercise session. Participants will complete a supervised bout of exercise on a treadmill 3x a week for 1-month. Participants will exercise at an RPE of 15-17 based on the participant's VO2peak test.	Behavioral: High Intensity Exercise; exercise at an intensity midway between the lactate threshold and VO2peak, from the screening test at an RPE of ~ 15-17; Dietary supplement: Nitrate-Depleted Placebo Beetroot Juice; Subjects will consume 1 bottle of nitrate-depleted beetroot juice 2x a day
Active Comparator: Nitrate-Rich Beetroot Juice + Moderate Intensity Exercise (MIE); Participants assigned to this group will be instructed to consume 1 bottle of nitrate-rich beetroot juice (~400mg of nitrate each) 2x a day (morning a night) starting 2-hours before the first exercise session. Participants will complete a supervised bout of exercise on a treadmill 3x a week for 1-month. Participants will exercise at an RPE of 10-12 based on the participant's VO2peak test.	Behavioral: Moderate Intensity Exercise; exercise at the RPE at LT associated an ~ RPE of 10-12; Dietary supplement: Nitrate-Rich Beetroot Juice; subjects will consume 1 bottle of nitrate-rich beetroot juice (~400mg of nitrate each) 2x a day; Other Names: Inorganic Nitrate; James White Beet It
Placebo Comparator: Nitrate-Depleted Placebo Beetroot Juice + Moderate Intensity Exercise (MIE); Participants assigned to this group will be instructed to consume 1 bottle of nitrate-depleted beetroot juice 2x a day (morning a night) starting 2-hours before the first exercise session. Participants will complete a supervised bout of exercise on a treadmill 3x a week for 1-month. Participants will exercise at an RPE of 10-12 based on the participant's VO2peak test.	Behavioral: Moderate Intensity Exercise; exercise at the RPE at LT associated an ~ RPE of 10-12; Dietary supplement: Nitrate-Depleted Placebo Beetroot Juice; Subjects will consume 1 bottle of nitrate-depleted beetroot juice 2x a day

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Vascular Endothelial Function (Flow-Mediated Dilation [FMD])	Participants will be placed in a supine position with their left forearm slightly extended and supinated with legs straight. The arteries will be imaged using a high-resolution 7.5MHz linear array transducer at rest, during 5 minutes of forearm occlusion via cuff inflation, and continuously for 3 minutes post-occlusion; an EKG trigger will be used to capture images during end-diastole of the cardiac cycle.	Baseline and after 1-month of exercise training
Cardiorespiratory Fitness	Peak aerobic capacity (VO2peak) will be assessed using a symptom-limited graded exercise test on a treadmill. An IV will be placed and lactate measured during rest and after the completion of every 3-minute stage to determine lactate threshold at VO2peak.	Baseline and after 1-month of exercise training

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Skeletal muscle perfusion	Skeletal Muscle Blood Flow (straight gauge plethysmography) will be measured non-invasively using venous occlusion/strain-gauge plethysmography both at rest and during reactive hyperemia following 5-minutes of occlusion via the Hokanson A16. A strain gauge device will be place around the widest part of the participant's right calf. A blood pressure place around participant's ankle will be inflated for 60 seconds to eliminate foot circulation from the measurement. At the same time, a cuff placed on the thigh will be inflated to block venous blood flow. The cuff occlusion will be held for ~60 seconds ((several cardiac cycles (4-6 cycles)) to obtain resting blood flow measurements.	Baseline and after 1-month of exercise training
Peripheral and Central Blood Pressures	Brachial artery blood pressures will be obtained using a standard sphygmomanometer. Aortic blood pressures will be obtained using applanation tonometry (SphygmoCor version 8.0, AtCor Medical).	Baseline and after 1-month of exercise training
24-hour ambulatory blood pressure	Blood pressure will be monitored using a standard ambulatory brachial artery blood pressure cuff (SunTech Oscar 2, SunTech Medical). Blood pressure measurements are programmed to be taken every 30-minutes during the day and every hour at night. This allows for monitoring of nighttime blood pressure changes and variations throughout the day.	Baseline and after 1-month of exercise training
Skeletal muscle microvascular perfusion (near-infra-red- spectroscopy)	Tissue oxygenation will be captured noninvasively using non-invasive, near-infrared spectrometry (NIRS, PortaMon, Artinis Medical Systems B.V., The Netherlands) positioned on the gastrocnemius or vastus lateralis muscle.	Baseline and after 1-month of exercise training
Hemoglobin	A blood sample will be obtained to measure hemoglobin.	Baseline and after 1-month of exercise training
Plasma Nitrate	A fasting blood draw will be taken. Following the blood draw, the participant will take one dose of Beetroot juice. The participant will then return to the lab three hours later for a second blood draw to assess level of nitrate in the plasma.	Baseline and after 1-month of exercise training
Plasma Nitrite	A fasting blood draw will be taken. Following the blood draw, the participant will take one dose of Beetroot juice. The participant will then return to the lab three hours later for a second blood draw to assess level of nitrite in the plasma.	Baseline and after 1-month of exercise training
Oral Nitrate Reducing Capacity	Assessment of the oral microbiome's ability to reduce nitrate to nitrite. This will be measured through an unstimulated saliva sample and a rinse of a standard nitrate solution.	Baseline and after 1-month of exercise training
Lipids	A blood sample will be obtained to measure lipids.	Baseline

Collaborators and Investigators

• Sponsor: University of Virginia
• Investigators: Principal Investigator: Jason Allen, PhD, UVA

Study record dates

Study Major Dates

• Study Start (Actual): July 1, 2025
• Primary Completion (Estimated): December 31, 2026
• Study Completion (Estimated): December 31, 2026

Study Registration Dates

• First Submitted: June 17, 2024
• First Submitted That Met QC Criteria: June 24, 2024
• First Posted (Actual): June 28, 2024

Study Record Updates

• Last Update Posted (Actual): July 28, 2025
• Last Update Submitted That Met QC Criteria: July 23, 2025
• Last Verified: July 1, 2025

More Information

Terms related to this study

• Keywords: Menopause; Vascular Health; Inorganic Nitrate; High Intensity Exercise
• Other Study ID Numbers: HSR231665

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No