Effect of Aronia Supplementation on the LPS-CD14-TLR2/4 Axis in Football Players

N/ANot Yet RecruitingNCT07829523

Hanna Dziewiecka40 enrolled

Overview

This randomized, double-blind, placebo-controlled trial will investigate the effects of 8-week Aronia melanocarpa supplementation on exercise-induced activation of the LPS-LBP-CD14-TLR2/TLR4 axis in trained male football players. Participants will be randomly assigned to receive either Aronia melanocarpa supplementation or placebo. Exercise testing will be performed before and after the 8-week intervention, with blood samples collected before, immediately after, and 1 hour after exercise. The study will assess circulating markers of endotoxemia and innate immune activation (LPS, LBP, sCD14, sTLR2, and sTLR4), TLR2 and TLR4 expression on monocyte subsets using flow cytometry, and TLR2 and TLR4 gene expression in peripheral blood mononuclear cells. The study aims to determine whether Aronia melanocarpa supplementation modulates the immune response to high-intensity intermittent exercise in football players.

High-intensity intermittent exercise, such as that performed during football, may induce gastrointestinal disturbances and transient alterations in intestinal barrier integrity. Exercise-related redistribution of blood flow away from the gastrointestinal tract, together with metabolic and thermal stress, may facilitate the translocation of bacterial products, including lipopolysaccharide (LPS), into the systemic circulation. LPS interacts with lipopolysaccharide-binding protein (LBP) and CD14 and may subsequently activate Toll-like receptors, particularly TLR4 and TLR2, contributing to the activation of innate immune responses. Aronia melanocarpa is a rich source of polyphenolic compounds, particularly anthocyanins and proanthocyanidins, with antioxidant and anti-inflammatory properties. The present study will investigate whether 8 weeks of Aronia melanocarpa supplementation can modulate exercise-induced responses within the LPS-LBP-CD14-TLR2/TLR4 axis in trained male football players. The study is designed as a randomized, double-blind, placebo-controlled trial. Thirty trained male football players aged 18-35 years will be recruited, allowing for an anticipated dropout rate, with a target sample size of 27 participants completing the study. Participants will be randomly assigned to an Aronia supplementation group or a placebo group. Randomization and coding of the capsules will be performed independently, and both participants and investigators will remain blinded to group allocation until completion of data collection and analysis. Participants assigned to the intervention group will receive three capsules per day for 8 weeks. Each capsule contains 150 mg of dry Aronia melanocarpa extract standardized to 18% anthocyanins. Participants assigned to the placebo group will receive visually identical capsules containing Aronia fiber and technological excipients. Participants will be instructed to take one capsule with each main meal. Exercise testing will be conducted before the start of supplementation and after the 8-week intervention. Exercise capacity will be assessed using a maximal multistage 20-m shuttle run (Beep Test). Before each exercise session, participants will follow standardized pre-test procedures, including avoidance of alcohol and intense physical exercise for 48 hours, avoidance of caffeine for 24 hours, and adherence to a standardized low-FODMAP diet during the 24 hours preceding testing. Venous blood samples will be collected before exercise, immediately after exercise, and 1 hour after exercise during both study sessions. The biological assessment will include circulating LPS, LBP, soluble CD14 (sCD14), soluble TLR2 (sTLR2), and soluble TLR4 (sTLR4). Flow cytometry will be used to characterize monocyte populations based on CD14 and CD16 expression and to assess TLR2 and TLR4 expression on these cells. In addition, TLR2 and TLR4 gene expression will be assessed in peripheral blood mononuclear cells using quantitative PCR. By combining circulating biomarkers with cellular and gene-expression measurements, the study will provide a multilevel assessment of the LPS-LBP-CD14-TLR2/TLR4 pathway. The primary objective is to determine whether 8-week Aronia melanocarpa supplementation modifies the exercise-induced activation of this pathway in trained football players.

Outcomes

Primary Outcomes

Exercise-Induced Change in Circulating Lipopolysaccharide (LPS) Concentration

Circulating lipopolysaccharide (LPS) concentration will be measured using an ELISA assay in blood samples collected before exercise, immediately after exercise, and 1 hour after exercise. Measurements will be performed during two study sessions: before the intervention and after 8 weeks of Aronia melanocarpa supplementation or placebo. The outcome will be used to assess exercise-induced changes in circulating LPS and whether these responses differ between the Aronia and placebo groups.