Effect of Chokeberry Supplementation on Oxidative Stress and Gut Integrity in Swimmers.

Effect of Chokeberry Supplementation on Oxidative Stress and Gut Microbiota in Swimmers.

This study included 8 weeks of supplementation of black chokeberry extract with 18% standardization of the anthocyanins content (dose 2×200 mg per day) or a placebo product made from chokeberry fiber. The first research data was collected among twenty-four homogeneous female swimmers who trained intensively and attended the Sports Championship School Complex. The second research data was collected among a group of twenty-two selective and homogeneous female swimmers who attended the Sports Championship School Complex extensively.

In this study, the athletes were subjected to intensive swimming tests in breaststroke crawl (800m + 200m + 50m) with 15 minutes of active rest between test sections in the water.

Blood samples were taken from the cubital vein at three time points: before stress test (pre-exercise), one minute after the end of the test (post-exercise), and after a 3-hour recovery period (3h recovery). Measurements of morphology, 4-Hydroxynonenal (4-HNE), 8-isoprostane, cortysol, Intestinal fatty acid binding protein (I-FABP), claudin3, lipopolysaccharide (LPS), lipopolysaccharide binding protein (LBP), and levels were determined using ELISA kits (SunRed Biotechnology Company).

Study Overview

• Status: Completed
• Conditions: Intestinal Permeability; Stress Oxidative
• Intervention / Treatment: Drug: Black chokeberry extract in capsules with 18% standardization of the anthocyanins content; Drug: Chokeberry fiber in capsules

Detailed Description

This study included 8 weeks of supplementation of black chokeberry extract with 18% standardization of the anthocyanins content (dose 2×200 mg per day), or a placebo product made from chokeberry fiber. The first date of research completed the group of twenty-four homogeneous female swimmers, who train swimming intensively and attend the Sports Championship School Complex. The second date of research completed the group of twenty-two selective and homogeneous female swimmers, who train swimming intensively and attend the Sports Championship School Complex.

In this study, the athletes were subjected to intensive swimming tests in breaststroke crawl (800m + 200m + 50m) with 15 minutes of active rest between test sections in the water. Blood samples were taken from the cubital vein at three time points: before stress test (pre-exercise), one minute after the end of the test (post-exercise), and after a 1-hour recovery period (3h recovery).

Measurement: All determined parameters were measured with the available equipment in the ZWKF laboratory in Gorzów Wielkopolski and using commercial assay kits. Measurements were performed by the project contractors.

1. Polyethylene tubes (2.7ml) containing dipotassium ethylenediaminetetraacetic acid (EDTA) anticoagulant were used for the following tests: (a) complete blood count (7 parameters) determined on the MYTHIC 18 hematology analyzer (Orphee Medical, Geneva, Switzerland). Red blood cell indices: RBC (Red Blood Cells), HGB (Hemoglobin), HCT (Hematocrit), MCV (Mean Corpuscular Volume), MCH (Mean Corpuscular Hemoglobin), MCHC (Mean Corpuscular Hemoglobin Concentration), RDW (Red Blood Cell Distribution Width); (b) a manual blood smear was made by placing a drop of blood on a slide and then spreading it with a uniform motion. After drying, it was colored by the May Grunwald-Giemsa method according to the procedure. After drying, the stained and fixed smear was viewed under a microscope for quantitative and qualitative assessment.
2. Polyethylene clotting activator tubes (9 ml) were centrifuged to separate the morphotic elements from the serum using a centrifuge (3000 rpm for 10 min). The serum was pipetted into several Eppendorf tubes, which were then frozen (temp. -80 °C). All biochemical parameters were determined from the extracted serum: (a) using the ELISA method by the test manufacturer's instructions. The designations include the following parameters:

4-Hydroxynonenal (4-HNE), 8-isoprostane, cortysol, Intestinal fatty acid binding protein (I-FABP), claudin3, lipopolysaccharide (LPS), lipopolysaccharide binding protein (LBP).

(3) The lactate (La) concentration was determined from the capillary blood immediately after collection using a commercially available kit (Dr. Lange, Germany).

A nutritionist thoroughly analyzed the athletes' diets before each exercise test. Using a commercially available program, the amount of energy, protein, carbohydrates, fats, and fiber was then analyzed.

The experiment was conducted in accordance with the Declaration of Helsinki. The Ethical Committee of the Medical University of Poznan approved the study protocol.

• Study Type: Interventional
• Enrollment (Actual): 22
• Phase: Phase 4

Contacts and Locations

Study Locations

• Poland
  • Poznań, Poland, 61-871
    • Poznań University of Physical Education

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child
• Accepts Healthy Volunteers: No

Description

Inclusion Criteria:

• valid medical examination,
• consent of the legal guardian of the underage athlete to participate in the study,
• Training experience of a minimum of three years
• Student of a sports championship school team,
• completion of the exercise test in the swimming pool - crawl style intensive test (800 m + 200 m + 50 m) on two test deadlines

Exclusion Criteria:

• antibiotic therapy,
• supplementation, use of any drugs, or oral hormonal contraceptives
• health problems within the last month

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Triple

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: supplemented; The supplemented group (n=12) consumed capsules with highly concentrated 18% chokeberry extract (one capsule contained 200 mg). Each capsule also contained: chokeberry fiber, E460b magnesium salts of stearic acid (anti-caking agent), E551 silicon dioxide (anti-caking agent), hydrocypropyl methylcellulose (capsule shell) and E171 titanium dioxide (shell colour). The producer of all capsules is MLD Biotrade in Poznan. The swimmers took 2 capsules a day (before breakfast and dinner) for nearly 56 days (depending on length menstrual cycle phase). The swimmers sipped each capsule with glass of water.	Drug: Black chokeberry extract in capsules with 18% standardization of the anthocyanins content; Supplement group: supplement of diet - black chokeberry extract (capsules) with 18% standardization of the anthocyanins content (one capsule contained 200 mg). Each capsule also contained: chokeberry fiber, E460b magnesium salts of stearic acid (anti-caking agent), E551 silicon dioxide (anti-caking agent), hydrocypropyl methylcellulose (capsule shell) and E171 titanium dioxide (shell colour). The producer of all capsules is MLD Biotrade in Poznan.
Placebo Comparator: Placebo; The control group (n=10) consumed capsules that were made from chokeberry fiber. The producer of all capsules is MLD Biotrade in Poznan. The swimmers took 2 capsules a day (before breakfast and dinner) for nearly 56 days (depending on length menstrual cycle phase). The swimmers sipped each capsule with glass of water.	Drug: Chokeberry fiber in capsules; Placebo group: placebo product - chokeberry fiber (placebo capsules, which were made from chokeberry fiber). The producer of all capsules is MLD Biotrade in Poznan.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Changes from baseline in 4-Hydroxynonenal (4-HNE) level.	Concentration of 4-Hydroxynonenal (4-HNE) [pg/ml]. ELISA method by the test manufacturer's instructions	At rest (before the test), directly after the test, and after a 3-hour recovery period.
Changes from baseline in 8-isoprostane level.	Concentration of 8-isoprostane [ng/mL]. ELISA method by the test manufacturer's instructions	At rest (before the test), directly after the test, and after a 3-hour recovery period.
Changes from baseline in Cortysol level.	Concentration of cortysol [ng/mL]. ELISA method by the test manufacturer's instructions	At rest (before the test), directly after the test, and after a 3-hour recovery period.
a manual blood smear	was made by placing a drop of blood on a slide and then spreading it with a uniform motion. After drying, it was colored by the May Grunwald-Giemsa method according to the procedure. The stained and fixed smear after drying was viewed under a microscope for quantitative and qualitative assessment.	At rest (before the test), directly after the test, and after a 3-hour recovery period.
complete blood count	Red blood cell indices: RBC (Red Blood Cells) [106 × µL-1], HGB (Hemoglobin) [g/dL] , HCT (Hematocrit) [%], MCV (Mean Corpuscular Volume)[fl], MCH (Mean Corpuscular Hemoglobin) [pg], MCHC (Mean Corpuscular Hemoglobin Concentration) [g x dL-1] , RDW (Red Blood cells Distribution Width) [%] .	At rest (before the test), directly after the test, and after a 3-hour recovery period.
Changes from baseline in Intestinal fatty acid binfing protein (I_FABP) level.	Concentration of I-FABP [pg/ml]. ELISA method by the test manufacturer's instructions	At rest (before the test), directly after the test, and after a 3-hour recovery period.
Changes from baseline in Claudin3 level.	Concentration of Claudin3 [ng/ml]. ELISA method by the test manufacturer's instructions	At rest (before the test), directly after the test, and after a 3-hour recovery period.
Changes from baseline in lipopolisacharide (LPS) level.	Concentration of LPS [EU/l]. ELISA method by the test manufacturer's instructions	At rest (before the test), directly after the test, and after a 3-hour recovery period.
Changes from baseline in lipopolisacharide binding protein (LBP) level.	Concentration of LBP [ug/ml]. ELISA method by the test manufacturer's instructions	At rest (before the test), directly after the test, and after a 3-hour recovery period.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Antropometric characteristic - height	Prior to the exercise test, the investigators measured the anthropometric parameters, including height (Seca 213 Hamburg, Deutschland) [cm]	Day 1 after overall fast
Antropometric characteristic - weight	Prior to the exercise test, the investigators measured the anthropometric parameters, including weight (Tanita BC 418 MA, Tanita Corporation, Tokyo, Japan) [kg]	Day 1 after overall fast
Food record - energy	Participants prepared a food record. The results were calculated using the dietetykpro program: energy [kcal]	Day before the Day 1, in the morning at Day 1
Food record - protein	Participants prepared a food record. The results were calculated using the dietetykpro program: protein [g]	Day before the Day 1, in the morning at Day 1
Food record - carobhydrates	Participants prepared a food record. The results were calculated using the dietetykpro program: carobhydrates [g]	Day before the Day 1, in the morning at Day 1
Food record - fiber	Participants prepared a food record. The results were calculated using the dietetykpro program: fiber [g]	Day before the Day 1, in the morning at Day 1
Food record - fat	Participants prepared a food record. The results were calculated using the dietetykpro program:: fat [g]	Day before the Day 1, in the morning at Day 1

Collaborators and Investigators

• Sponsor: Poznan University of Physical Education

Study record dates

Study Major Dates

• Study Start (Actual): March 1, 2023
• Primary Completion (Actual): December 28, 2023
• Study Completion (Actual): December 28, 2023

Study Registration Dates

• First Submitted: March 24, 2025
• First Submitted That Met QC Criteria: March 24, 2025
• First Posted (Actual): March 30, 2025

Study Record Updates

• Last Update Posted (Estimated): July 8, 2025
• Last Update Submitted That Met QC Criteria: July 2, 2025
• Last Verified: March 1, 2025

More Information

Terms related to this study

• Other Study ID Numbers: SWIMMER- SUPL

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
• product manufactured in and exported from the U.S.: No