Effects of Probiotics on Gut Microbiota, Endocannabinoid and Immune Activation and Symptoms of Fatigue in Dancers

February 9, 2026 updated by: Jakub Wiącek, Poznan University of Physical Education

Effects of 3-month Probiotic Mix Supplementation (L. Helveticus R-0052, B. Longum R-0175) on Gut Microbiota and Metabolome, Endocannabinoid and Immune Systems Activation, Along With Symptoms of Fatigue in Professional Dancers

The aim of the planned research is to assess the dynamics of changes in the elements of the gut-brain axis (GBA), the cytokine profile and the endocannabinoid system markers, after dietary supplementation with probiotics Lactobacillus helveticus Rosell-52 and Bifidobacterium longum Rosell-175 by professional dancers. Although in recent years there has been growing interest in the influence of the gut microbiota on the body's adaptation to stress stimuli and on overall health, there is a lack of information on the influence of probiotics on systems involved in maintaining neuropsychiatric balance, such as the endocannabinoid system.

In order to determine the validity of the applied therapy with selective probiotics, the following will be assessed: intestinal bacteria and bacterial metabolites in the stool, cannabinoids and cannabinoid receptors and enzymes in the blood, indicators of mental distress in the blood, cytokines responsible for the modulation of the gut-brain axis in the blood, as well as questionnaires regarding the functioning of the digestive tract, fatigue, stress and sleep quality.

The study will involve active dancers of the Polish Theater in Poznan, the Polish Dance Theater, the Private School of Dance Art in Poznan and students of the Academy of Physical Education in the field of Dance. Dancers are a group of athletes that is exposed to particular injuries and work-overload. Professional dancers spend multiple hours a week on intensive physical training. The largest percentage of injuries occurring in the group of professional dancers are chronic injuries, including: inflammation of soft tissues, muscle strains and tears.

Professional dance is one of the most physically demanding forms of physical activity, and at the same time it is associated with a high burden on the nervous system problems caused by performances in front of an audience or subjective jury, frequent traveling and disturbances in the circadian rhythm.

Study Overview

Status

Completed

Conditions

Intervention / Treatment

Detailed Description

Professional dancers are often referred to as artistic athletes due to their high level of physical activity and competition. Training dance for over 11.5 hours a week increases the risk of injury (women - mostly bone injuries, men - mostly bruises and tendon injuries). One of the reasons for the increased susceptibility to injuries among dancers is psychological stress. The observations results indicate that dancers often decide to continue training despite feeling pain, which limits the smoothness of their movements and leads to a further increase of stress. Highly physically active groups of people are more exposed to functional dyspepsia (FD) and irritable bowel syndrome (IBS), which occurence is correlated to chronic stress. As a result of chronic stress and the activation of the hypothalamic pituitary-adrenal axis (HPA), inflammatory processes in the gastrointestinal mucosa are initiated. The presence of noradrenaline increases the adhesion of bacteria and viruses to the epithelium, which in turn modulates the immune system within the intestines and increases their inflammation. Systemically acting cortisol causes the leakage of tight junctions (TJ) cells and an increase in the permeability of the intestinal barrier. Inflammation stimulates the secretion of cortisol, which initiates the further cycle of inflammation and weakening of the intestinal barrier function. As a consequence, cytokines and inflammatory mediators released in the course of inflammation act directly on the nerve endings that transmit these afferent nerve signals to the brain, becoming an endogenous stressor.

When exogenous and endogenous stress factors overlap, the body's perception of stress increases, which ultimately leads to psychological, physiological and behavioral changes.

Pain and depression are common comorbidities of neuropsychiatric origin. One of the body's regulatory systems, which in both cases is dysfunctional, is the endocannabinoid system (ECS). It has been suggested that these diseases can be caused by intense and prolonged exercise, which disrupts the intestinal barrier, causing changes in the profile of metabolites and the function of the intestinal microbiota. Overtraining leads to an increase in the levels of cortisol, adrenaline and noradrenaline, and to the translocation of lipopolysaccharide (LPS) outside the gut, increasing the concentration of pro-inflammatory cytokines in the body. This results in dysregulation of the balance between serotonin (5-HT), dopamine (DA) and gamma-aminobutyric acid (GABA) and fatigue. These changes lead to further activation of the endocannabinoid system and, in the long-term, weakening of adaptive abilities.

The neuromodulatory properties of the cannabinoid system are manifested e.g. in short- and long-term synaptic plasticity and modulation of pain conduction. Research results indicate that the endocannabinoid system, in addition to its key role in regulating intestinal motility, also affects the secretory functions of the gastrointestinal tract and the integrity of the intestinal epithelium, which may be an alternative way to regulate the immune system and inflammation in the intestines.

Type 1 cannabinoid receptors (CB1) are located presynaptically in the cell membrane of central and peripheral nervous system neurons, and their activation inhibits the release of many neurotransmitters, i.e. acetylcholine, noradrenaline, dopamine, serotonin, glutamate and γ-aminobutyric acid. Type 2 cannabinoid receptors (CB2) are found mainly on the surface of cells of the immune system, especially B-lymphocytes, macrophages and monocytes. Their activation inhibits pro-inflammatory cytokines release and increases the release of anti-inflammatory cytokines.

Cannabinoid receptors are found in both, the immune and digestive systems. Endogenous cannabinoids modify the body's response to stress by influencing the hypothalamic pituitary-adrenal axis (HPA). Stress leads to an increased activity of endocannabinoids which, mainly through the CB1 cannabinoid receptors, lead to the inhibition of the release of corticosteroids. In addition, the activation of CB1 receptors in the gastrointestinal tract reduces the intensity of pain conduction (nociception), which is induced by activation, e.g. TRPV1 vanilloid receptors. It has been shown that inflammation within the intestinal epithelium of the gastrointestinal tract causes an increase in the neural conduction of neurons containing CB1 receptors, which contributes to a change in the proportion of these receptors in relation to the TRPV1 located mainly in the cell membrane of dorsal root ganglion afferents (DRGs). As a result, two different receptors interact and the TRPV1 receptor activity is abolished. Inflammatory mediators released from the intestinal epithelium stimulate visceral pain, while activation of CB2 receptors probably reduces their action.

There are indications that a qualitative and quantitative change in the intestinal microbiota may affect the activity of endogenous ligands and mediators of the endocannabinoid system (ECS). Although the mechanisms of regulation of the level of endocannabinoids and related bioactive lipids by selected bacteria are not fully understood, it has been shown that probiotic supplementation can induce an increase in the concentration of endogenous cannabinoids, i.e. 2-AG (2-arachidonoylglycerol), 2-OG (2-oleoylglycerol) and 2-PG (2-palmitoylglycerol). In an animal experiment, the use of monoacylglycerol lipase inhibition reduced the degradation of 2-AG, which reduced endotoxemia and systemic inflammation. In another study, the deletion of the Myd88 gene encoding the TLR (toll-like receptor) receptor protein in intestinal epithelial cells changed the composition of the intestinal microbiota, decreased the synthesis of anandamide (AEA) and increased the synthesis of anti-inflammatory endocannabinoids, such as: 2-AG, 2-PG, 2 OG. Released endocannabinoids with anti-inflammatory properties have the ability to activate so-called orphan GPR119 (G protein-coupled receptor 119) receptors, which are associated with the secretion of anti-inflammatory mediators. Moreover, it has been shown that an increase in LPS concentration induces AEA synthesis and a decrease in fatty-acid amide hydrolase (FAAH) in macrophages, as well as an increase in AEA production in peripheral lymphocytes, which may be important for the regulation of the intestinal barrier function and the level of inflammation. The results of studies carried out on a mouse model revealed the importance of the intestinal microbiota in the regulation of the expression of the NAPE-PLD gene (phospholipase D specific for n-acylphosphatidylethanolamine), i.e. an enzyme involved in the synthesis of anandamide and in the selective regulation of CB1 mRNA (messenger ribonucleic acid) expression. In an obese mouse model, it was observed that the administration of a probiotic decreased the expression level of CB1 receptor mRNA while reducing the concentration of AEA ligand and increasing the expression of FAAH mRNA. In other studies, administration of a probiotic to mice resulted in a decrease in the concentration of LPS in the blood plasma, which correlated with both the level of AEA and the expression of CB1 mRNA in the colon epithelium. In in vitro and in vivo experiments on animal models, it has been observed that CB2 receptors are activated when an imbalance in the innate immune system occurs. The NLRP3 inflammasomes are suppressed by autophagy, a mechanism that may be involved in the suppression of inflammation and the regulation of the intestinal barrier, e.g. in irritable bowel syndrome. Another factor that is a potential NLRP3 inhibitor is the intestinal microbiota metabolite, butyrate, which belongs to the short-chain fatty acids. Its action has a positive effect on the functions of the intestinal epithelial cells and the stability of the intestinal barrier. In studies conducted on germ-free mice, after the intestinal microbiota was transferred, the metabolism of endocannabinoids in the gastrointestinal tract changed. The use of probiotic therapy in mice with previously induced intestinal dysbiosis changed the activation of endocannabinoid receptors, the behavior of animals and a decrease in intestinal inflammation. Other research results indicate that inhibition of the CB1 receptor in obese mice stimulates the secretion of mucin, which is the primary source of nutrients for the development of Akkermansia muciniphila. This observation confirms the results of studies by other authors, who demonstrated the possibility of regulation of CB2 receptors by some species of intestinal bacteria and the related immune response.

The mechanisms of the transmission of inflammatory signals from the gut to the CNS (central nervous system) are not fully elucidated. The results of the research indicate, however, that the systemic inflammation accompanying depressive symptoms is associated with changes in the ecosystem of the intestinal microbiota and the production of SCFA (short-chain fatty acids) and other metabolites of a neurobiological nature. Moreover, it can also lead to changes in the synthesis and release of endocannabinoids or the metabolic pathways of tryptophan and kynurenine (KYN). So far, it has been observed that there is a link between ECS and the kynurenine pathway in neurological disorders such as epilepsy or migraine headaches, which are associated with excessive cell stimulation and excitotoxicity. The latest research results indicate the possibility of modifying the transformations of tryptophan and kynurenine by probiotic therapy.

There is limited number of human research on the role of the gut microbiota in the modulation of the ECS. The available literature lacks studies describing the relationship of changes in the intestinal microbiota and its metabolome under the influence of targeted, multi-strain probiotic therapy on the response of the endocannabinoid and immune system in people subjected to high physical and mental stress. Observations that will be made in dancers supplemented with a multi strain probiotic containing Lactobacillus helveticus Rosell-52 and Bifidobacterium longum Rosell-175 may help finding effective tools in the treatment of gastrointestinal disorders and stress related to it. A potential mechanism behind this action may be the restoration of the normal gut microbiota and the profile of its metabolites, as well as the improvement of the gut barrier and endocannabinoid function.

Study Type

Interventional

Enrollment (Actual)

26

Phase

  • Not Applicable

Contacts and Locations

This section provides the contact details for those conducting the study, and information on where this study is being conducted.

Study Locations

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

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Ages Eligible for Study

14 years to 32 years (Adult)

Accepts Healthy Volunteers

Yes

Description

Inclusion Criteria:

  • Age > 18, <36 years old;
  • Professional dancing activity with over 8 hours of training per week.

Exclusion Criteria:

  • Age <18,> 36 y.o.;
  • Being injured within 3 months from the start of the study;
  • Taking pre- and / or probiotics in the last 3 months before the study;
  • Hospitalization during the last 4 weeks before starting of the study;
  • Traveling to tropical countries during the last 4 weeks before study;
  • Taking antibiotics, steroids and anabolic steroids in the last 4 weeks before study.

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

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

Arms and Interventions

Participant Group / Arm
Intervention / Treatment
Experimental: Probiotic treatment group
ca. 30 participants
Dietary supplement: Lactobacillus helveticus Rosell-52 and Bifidobacterium longum Rosell-175 (Sanprobi Stress)
3-month, multi-strain probiotic supplementation (one a day, 3 x 10⁹ CFU)
Other Names:
  • L. helveticus R-0052, B. longum R-0175
Placebo Comparator: Placebo group
ca. 30 participants
Dietary supplement: Starch (placebo)
3-month placebo
Other Names:
  • capsules containing starch

What is the study measuring?

Primary Outcome Measures

Outcome Measure
Measure Description
Time Frame
Qualitative Assessment of Intestinal Microbiota in Stool Samples
Time Frame: Baseline and 3 months
• gut bacteria species in stool will be assessed using the shallow shotgun sequencing method, NGS (Next-Generation Sequencing);
Baseline and 3 months
Quantitive Change in Intestinal Microbiota in Stool Samples
Time Frame: Baseline and 3 months
• determination of quantitative (Colony forming units - CFU) changes in bacteria in the stool - the shallow shotgun sequencing method, NGS (Next Generation Sequencing) molecular analysis will be used to assess the changes;
Baseline and 3 months
Intestinal Metabolome in Stool Samples
Time Frame: Baseline and 3 months
Metabolites with fold change >1.5 and p<0.05 were considered significantly altered. The reported values represent the number of metabolites meeting these criteria in the probiotic group. • metabolomic analysis (non-targeted metabolome, short-chain fatty acids, trimethylamines, tryptophan catabolites) will be performed on a quadrupole mass spectrometer coupled with a time-of-flight (QToF) analyzer connected to the high performance liquid chromatograph (UHPLC).
Baseline and 3 months
Quantitive Change in Endocannabinoids Levels in Blood Samples
Time Frame: Baseline and 3 months
• determination of endocannabinoids and cannabinoid receptors: anandamide (AEA) using ELISA Kit (nanograms per millilitre (ng/mL));
Baseline and 3 months
Quantitive Change in Endocannabinoids' Receptors Levels in Blood Samples
Time Frame: Baseline and 3 months
• determination of cannabinoid receptors: Endocannabinoid Receptor 2 (CNR2) using ELISA Kit (nanograms per millilitre (ng/mL));
Baseline and 3 months
Quantitive Change in Endocannabinoids Metabolism Enzymes in Blood
Time Frame: Baseline and 3 months
• determination of cannabinoid metabolism enzymes: fatty acid amide hydrolase (FAAH) using ELISA Kit (nanograms per millilitre (ng/mL));
Baseline and 3 months
Quantitive Change in Intestinal Barrier Proteins in Blood Samples
Time Frame: Baseline and 3 months
• determination of blood biomarkers of a disturbed intestinal barrier concentrations: zonulin, calprotectin (CALPRO) using ELISA Kit (nanograms per millilitre (ng/mL));
Baseline and 3 months
Quantitive Change in Intestinal Barrier Biomarkers in Blood Samples
Time Frame: Baseline and 3 months
• determination of blood biomarkers of a disturbed intestinal barrier concentrations: lipopolysaccharide (LPS), using ELISA Kit (picograms per millilitre (pg/mL));
Baseline and 3 months
Quantitive Change in Tumor Necrosis Factor-alpha (TNF-α) in Blood Samples
Time Frame: Baseline and 3 months
• determination of blood tumor necrosis factor-alpha (TNF-α) concentration using ELISA Kit (nanograms per millilitre (ng/mL));
Baseline and 3 months
Quantitive Change in Leukemia Inhibitory Factor (LIF) in Blood Samples
Time Frame: Baseline and 3 months
• determination of blood leukemia inhibitory factor (LIF) concentration using ELISA Kit (nanograms per millilitre (ng/mL));
Baseline and 3 months
Quantitive Change in Blood Interleukins Profile
Time Frame: Baseline and 3 months
• determination of interleukins concentrations: IL-1β (interleukin-1beta), using ELISA Kit (picograms per millilitre (pg/mL));
Baseline and 3 months
Quantitive Change in Chronic Stress Biomarker in Blood Samples - Cortisol
Time Frame: Baseline and 3 months
• determination of blood cortisol concentration using ELISA Kit (nanograms per millilitre (ng/mL)) - Instead of cortisol, IL-10 levels were analyzed as a marker of anti-inflammatory response [pg/mL].
Baseline and 3 months
Quantitive Change in Inflammation Biomarker in Blood Samples - C-reactive Protein
Time Frame: Baseline and 3 months
• determination of C-reactive protein (CRP) concentration using ELISA Kit (picograms per millilitre (pg/mL));
Baseline and 3 months

Secondary Outcome Measures

Outcome Measure
Measure Description
Time Frame
Coping With Stress Questionnaire
Time Frame: Baseline and 3 months
• determination of methods of active coping with stress using the Inventory for Measuring Coping with Stress (Mini-COPE); It is designed to establish how a study participant behaves when experiences particular events; The scale: 0 = "I hardly ever do this", 1 = "I rarely do this", 2 = "I do this often", 3 = "I almost always do this"; [0-18] - higher scores mean better outcome
Baseline and 3 months
Fatigue Questionnaire
Time Frame: Baseline and 3 months
• assessment of the level of fatigue using the Fatigue Assessment Scale (FAS); the statements of the questionnaire relate to the feeling of well-being; The scale: 1. Never (0 points), 2. Sometimes (once a month or less) (1 point), 3. Regularly (several times a month) (2 points), 4. Often (weekly)(3 points) and 5. Always (everyday) (4 points); [0-32] - higher score means worse outcome
Baseline and 3 months
Gastrointestinal Disorders Questionnaire
Time Frame: Baseline and 3 months
• assessment of gastrointestinal pain using the Rome IV Questionnaire for adults (selected questions on irritable bowel syndrome, constipation and diarrhea); the statements of the questionnaire relate to frequency and intensity of disorders; [0-10] - higher scores mean worse outcome;
Baseline and 3 months
Sleep Quality Questionnaire
Time Frame: Baseline and 3 months
• assessment of the sleep quality level using Pittsburgh Sleep Quality Questionnaire (PSQI); the statements of the questionnaire relate to frequency of sleep disorders; [0-3] - higher scores mean worse outcome;
Baseline and 3 months
Body Mass Analysis (BMI, kg/m^2)
Time Frame: Baseline and 3 months
• determination of body weight and composition using the electrical bioimpedance method;
Baseline and 3 months
Body Composition Analysis (Percent of Total Body Mass)
Time Frame: Baseline and 3 months
• assessment of bone, fat and fat-free mass using the electrical bioimpedance method;
Baseline and 3 months
Composite Measure of Diet Composition Assessed by Food Diaries
Time Frame: Baseline and 3 months
• assessment of protein, fat, carbohydrates and fibre intake using food diaries (grams);
Baseline and 3 months
Pain Threshold Test (Newtons)
Time Frame: Baseline and 3 months
• mechanical stimuli pain threshold assessment using an algometer. Mechanical pressure pain threshold was assessed on the thumb flexor muscle using a pressure algometer. The probe was applied perpendicularly with gradually increasing pressure until the participant indicated the first painful sensation. Three consecutive measurements were performed, and the mean value in Newtons was recorded as the outcome.
Baseline and 3 months
Changes in Red Blood Cells Count
Time Frame: Baseline and 3 months
• determination of red blood cell counts using flow cytometry (trillion cells per Litre);
Baseline and 3 months
Changes in White Blood Cells Count
Time Frame: Baseline and 3 months
• determination of white blood cells counts using flow cytometry (billion cells per Litre);
Baseline and 3 months
Changes in Hemoglobin Level
Time Frame: Baseline and 3 months
• determination of hemoglobin level using flow cytometry (grams per Litre);
Baseline and 3 months
Changes in Hematocrit
Time Frame: Baseline and 3 months
• determination of hematocrit using flow cytometry (percentage of red blood cells in blood);
Baseline and 3 months
Changes in Platelets Counts
Time Frame: Baseline and 3 months
• determination of platelets counts using flow cytometry (billions per Litre)
Baseline and 3 months

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Sponsor

Poznan University of Physical Education

Investigators

  • Study Chair: Joanna Karolkiewicz, Professor, Poznan University of Physical Education

Publications and helpful links

The person responsible for entering information about the study voluntarily provides these publications. These may be about anything related to the study.

General Publications

Study record dates

These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.

Study Major Dates

Study Start (Actual)

October 1, 2022

Primary Completion (Actual)

March 8, 2023

Study Completion (Actual)

June 13, 2023

Study Registration Dates

First Submitted

September 22, 2022

First Submitted That Met QC Criteria

October 4, 2022

First Posted (Actual)

October 5, 2022

Study Record Updates

Last Update Posted (Actual)

February 20, 2026

Last Update Submitted That Met QC Criteria

February 9, 2026

Last Verified

February 1, 2026

More Information

Terms related to this study

Keywords

Additional Relevant MeSH Terms

Other Study ID Numbers

  • 11/NSN/4/2022

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

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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