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IRCT20260722070298N1 IRCT 2026-10-15 Oroumia University of Medical Sciences Effect of Synbiotic in MASLD Effect of Synbiotic Supplementation on Liver Fibrosis Indices, Inflammatory and Oxidative Stress Biomarkers, Appetite, Metabolic Parameters, and Expression of PPAR-α, UCP1, and UCP2 Genes in Overweight or Obese Adolescents with Metabolic Dysfunction–Associated Steatotic Liver Disease (MASLD): A Randomized Double-Blind Placebo-Controlled Trial 2026-10-22 anticipated 126 Recruiting https://irct.ir/trial/91817 interventional Randomization: Randomized, Blinding: Double blinded, Placebo: Used, Assignment: Parallel, Purpose: Treatment, Other design features: Standardized Lifestyle and Dietary Counseling:To isolate the specific effect of the synbiotic supplementation and control for potential confounding factors, both the intervention and placebo groups will receive identical, standardized dietary and lifestyle counseling throughout the 16-week trial. This counseling will be provided by a trained clinical nutritionist to both the adolescent and their parent (preferably the mother) at baseline and reinforced during follow-up visits.The dietary plan is designed based on established guidelines for the staged treatment of pediatric obesity, with the goal of achieving safe, gradual weight loss and improving metabolic parameters. The daily energy deficit is calculated individually based on the severity of obesity: For overweight adolescents (BMI-for-age between 85th and 95th percentile): A diet with a daily energy deficit of 300 kcal relative to their estimated energy requirement (EER). For obese adolescents (BMI-for-age ≥ 95th percentile): A diet with a daily energy deficit of 500 kcal relative to their EER.This approach aims for a gradual, safe, and sustainable weight loss of 0.5 to 1 kg per week. The dietary composition will follow standard recommendations for pediatric MASLD: Carbohydrates: 50-55% of total daily energy (with emphasis on complex, high-fiber sources like whole grains, vegetables, and legumes). Protein: 15-20% of total daily energy (from lean sources such as skinless poultry, fish, legumes, and low-fat dairy). Fat: 25-30% of total daily energy (with emphasis on unsaturated fats like olive oil, nuts, and seeds, while limiting saturated and trans fats).Behavioral and Dietary Recommendations: Complete elimination of sugar-sweetened beverages. Increased consumption of fresh fruits and vegetables to at least 5 servings per day. Controlled portion sizes and avoidance of fast food. Encouragement of at least 5 family meals per week. Self-monitoring of daily food intake using a provided diary.Monitoring Dietary Intake:To ensure that any observed changes in metabolic and inflammatory markers are primarily due to the synbiotic intervention and not unintended dietary changes, dietary intake will be rigorously monitored at three time points (baseline, week 8, and week 16) using three 24-hour dietary recalls at each time point. This data will be analyzed using Nutritionist IV software and will be included as a covariate in the final statistical analysis. Physical activity will also be assessed at baseline and week 16 using the validated Persian version of the Physical Activity Questionnaire for Adolescents (PAQ-A) and adjusted for in the analysis. This comprehensive approach ensures that the true effect of the synbiotic can be isolated from lifestyle modifications, Randomization description: This is a single-center, randomized, double-blind, placebo-controlled trial conducted at a clinical center affiliated with Urmia University of Medical Sciences. Eligible participants will be randomly allocated in a 1:1 ratio to either the synbiotic supplementation or placebo group. To ensure a balanced distribution of baseline confounding factors across both arms, stratified block randomization will be performed based on four key variables: sex (female vs. male), age (13–15 vs. 16–18 years), body mass index (<95th percentile vs. ≥95th percentile), and baseline Controlled Attenuation Parameter (CAP score: <260 dB/m vs. ≥260 dB/m), yielding a total of 16 distinct strata. For each stratum, an independent randomization sequence will be generated using STATA software (version 17) with the blockrand package and a fixed block size of 4, applying a predefined random seed to ensure reproducibility. Allocation concealment will be strictly maintained using sequentially numbered, opaque, sealed envelopes (SNOSE) prepared separately for each stratum. The envelopes and uniquely coded, identical-looking supplement/placebo containers will be prepared and managed by an independent researcher/pharmacist who has no involvement in participant recruitment, clinical care, or outcome assessment. Envelopes will only be opened sequentially after formal participant enrollment, Blinding description: The trial is double-blind. Participants, parents, investigators, clinicians/care providers, study personnel involved in outcome assessment, and data analysts will remain unaware of group assignments. Synbiotic and placebo capsules will be identical in appearance, color, smell, and taste. Capsule containers will be coded according to the allocation list by a pharmacist who will not be involved in data collection or analysis. The randomization code will be broken only after data collection or when potentially required for a serious adverse event. In the event of a serious adverse event, the need for unblinding will be jointly reviewed by the Principal Investigator and the clinical hepatologist. N/A Metabolic dysfunction-Associated Steatotic Liver Disease (MASLD) / Non-Alcoholic Fatty Liver Disease (NAFLD) in adolescents. Intervention 1: Intervention group: Participants will receive one capsule of Familact 2Plus daily (produced by Zist-Takhmir Co., Tehran, Iran). Each capsule contains 8 probiotic strains (Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus salivarius, Lactobacillus reuteri, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium bifidum) with a total of approximately 8×10⁹ CFU, and 38.5 mg of Fructooligosaccharides (FOS) as a prebiotic. The capsules should be taken on an empty stomach, approximately one hour before lunch, with a glass of water, for a duration of 16 weeks. Storage conditions: keep at temperatures below 25°C, in a dry place, away from light and moisture. In addition to the supplement, participants in this group will receive standardized dietary and lifestyle counseling. The dietary plan is designed based on established guidelines for the staged treatment of pediatric obesity, with the goal of achieving safe, gradual weight loss and improving metabolic parameters. The daily energy deficit is calculated individually based on the severity of obesity: For overweight adolescents (BMI-for-age between 85th and 95th percentile): A diet with a daily energy deficit of 300 kcal relative to their estimated energy requirement (EER). For obese adolescents (BMI-for-age ≥ 95th percentile): A diet with a daily energy deficit of 500 kcal relative to their EER.This approach aims for a gradual, safe, and sustainable weight loss of 0.5 to 1 kg per week. The dietary composition will follow standard recommendations for pediatric MASLD (50-55% carbohydrates, 15-20% protein, and 25-30% fat). Behavioral recommendations include elimination of sugar-sweetened beverages, increased consumption of fruits and vegetables (≥5 servings/day), controlled portion sizes, avoidance of fast food, encouragement of family meals, and self-monitoring of food intake using a diary. Intervention 2: Control group: Participants will receive a placebo capsule, visually identical to the intervention capsule, produced by Zist-Takhmir Co. The placebo capsules do not contain any active bacterial strains or prebiotic content. The dosage and administration schedule will be identical to the intervention group: one capsule daily, on an empty stomach, approximately one hour before lunch, with a glass of water, for a duration of 16 weeks. Storage conditions: keep at temperatures below 25°C, in a dry place, away from light and moisture.Participants in this group will receive standardized nutritional and lifestyle counseling. No - There is not a plan to make this available Justification or reason for not sharing IPD is The participants are adolescents, and the dataset contains sensitive clinical, biochemical, anthropometric, and gene-expression data. The informed consent and the current protocol do not include provisions for sharing individual participant data with external researchers. Therefore, to protect participant confidentiality and minimize the risk of re-identification, deidentified individual participant data will not be shared. Study findings will be disseminated only in aggregate form through scientific publications and presentations.
public Rasoul Zarrin
Urmia University of Medical Sciences, Nazlou Pardis, Sero Road, Urmia, Iran.
Urmia Iran (Islamic Republic of) 5756115198 +98 914 884 3592 wwzarrin@gmail.com Oroumia University of Medical Sciences scientific Rasoul Zarrin
azlou Pardis, Sero Road, School of Medicine, Urmia University of Medical Sciences, Urmia, Iran
Urmia Iran (Islamic Republic of) 5756115198 +98 914 884 3592 wwzarrin@gmail.com Oroumia University of Medical Sciences Iran (Islamic Republic of) Age 13 to 18 years. Overweight or obesity, defined as BMI-for-age ≥85th percentile according to WHO growth reference. Diagnosis of MASLD confirmed by FibroScan with CAP ≥248 dB/m. Stable dietary and physical activity patterns during the 3 months prior to enrollment. Written informed consent from the parent or legal guardian and assent from the participant. 13 years 18 years Both History of cardiovascular, renal, or autoimmune diseases. Use of hepatotoxic medications within 3 months before the study. Participation in another clinical trial within the previous 3 months. History of probiotic, prebiotic, or antibiotic supplement use within the past 3 months. Use of medications affecting metabolism, such as statins, glucocorticoids, metformin, or other insulin sensitizers. Presence of chronic liver diseases, including viral hepatitis, Wilson’s disease, autoimmune hepatitis, or genetic and metabolic liver disorders. Diagnosis of renal disease, thyroid disorders, or other endocrine abnormalities. Pregnancy at baseline in post-menarcheal female participants. K76.0 Fatty (change of) liver, not elsewhere classified Treatment - Other Placebo Intervention group: Participants will receive one capsule of Familact 2Plus daily (produced by Zist-Takhmir Co., Tehran, Iran). Each capsule contains 8 probiotic strains (Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus salivarius, Lactobacillus reuteri, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium bifidum) with a total of approximately 8×10⁹ CFU, and 38.5 mg of Fructooligosaccharides (FOS) as a prebiotic. The capsules should be taken on an empty stomach, approximately one hour before lunch, with a glass of water, for a duration of 16 weeks. Storage conditions: keep at temperatures below 25°C, in a dry place, away from light and moisture. In addition to the supplement, participants in this group will receive standardized dietary and lifestyle counseling. The dietary plan is designed based on established guidelines for the staged treatment of pediatric obesity, with the goal of achieving safe, gradual weight loss and improving metabolic parameters. The daily energy deficit is calculated individually based on the severity of obesity: For overweight adolescents (BMI-for-age between 85th and 95th percentile): A diet with a daily energy deficit of 300 kcal relative to their estimated energy requirement (EER). For obese adolescents (BMI-for-age ≥ 95th percentile): A diet with a daily energy deficit of 500 kcal relative to their EER.This approach aims for a gradual, safe, and sustainable weight loss of 0.5 to 1 kg per week. The dietary composition will follow standard recommendations for pediatric MASLD (50-55% carbohydrates, 15-20% protein, and 25-30% fat). Behavioral recommendations include elimination of sugar-sweetened beverages, increased consumption of fruits and vegetables (≥5 servings/day), controlled portion sizes, avoidance of fast food, encouragement of family meals, and self-monitoring of food intake using a diary. Control group: Participants will receive a placebo capsule, visually identical to the intervention capsule, produced by Zist-Takhmir Co. The placebo capsules do not contain any active bacterial strains or prebiotic content. The dosage and administration schedule will be identical to the intervention group: one capsule daily, on an empty stomach, approximately one hour before lunch, with a glass of water, for a duration of 16 weeks. Storage conditions: keep at temperatures below 25°C, in a dry place, away from light and moisture.Participants in this group will receive standardized nutritional and lifestyle counseling. Change in controlled attenuation parameter measured by FibroScan from baseline to week 16 (end of the intervention). Timepoint: At baseline and at week 16 (end of the intervention). Method of measurement: Controlled attenuation parameter will be measured using FibroScan transient elastography. Expression of Peroxisome Proliferator-Activated Receptor Alpha in peripheral blood mononuclear cells. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Quantitative real-time polymerase chain reaction using Beta-actin as the reference gene and the 2 to the power of negative delta delta Ct method. Expression of Uncoupling Protein 1 in peripheral blood mononuclear cells. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Quantitative real-time polymerase chain reaction using Beta-actin as the reference gene and the 2 to the power of negative delta delta Ct method. Expression of Uncoupling Protein 2 in peripheral blood mononuclear cells. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Quantitative real-time polymerase chain reaction using Beta-actin as the reference gene and the 2 to the power of negative delta delta Ct method. Alanine aminotransferase. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzymatic colorimetric method using a commercial kit and calibrated autoanalyzer. Aspartate aminotransferase. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzymatic colorimetric method using a commercial kit and calibrated autoanalyzer. Gamma glutamyl transferase. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzymatic colorimetric method using a commercial kit and calibrated autoanalyzer. Fasting blood glucose. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzymatic colorimetric method using a commercial kit and calibrated autoanalyzer after 12 hours of fasting. Fasting insulin. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzyme linked immunosorbent assay after 12 hours of fasting. Homeostatic Model Assessment of Insulin Resistance. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Calculated from fasting blood glucose and fasting insulin. Triglyceride. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzymatic colorimetric method using a commercial kit and calibrated autoanalyzer. Low density lipoprotein cholesterol. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzymatic colorimetric method using a commercial kit and calibrated autoanalyzer. High density lipoprotein cholesterol. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzymatic colorimetric method using a commercial kit and calibrated autoanalyzer. Total cholesterol. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzymatic colorimetric method using a commercial kit and calibrated autoanalyzer. Neutrophil to lymphocyte ratio. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Calculated from complete blood count. Platelet to lymphocyte ratio. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Calculated from complete blood count. C reactive protein. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzyme linked immunosorbent assay. Tumor necrosis factor alpha. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Enzyme linked immunosorbent assay. Erythrocyte sedimentation rate. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Standard clinical laboratory method. Malondialdehyde. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Thiobarbituric acid reactive substances assay. Superoxide dismutase activity. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Commercial colorimetric assay. Body weight. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Calibrated digital scale. Height. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Stadiometer. Body mass index. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Calculated from body weight and height. Waist circumference. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Non stretchable measuring tape. Conicity index. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Calculated from waist circumference, height, and body weight. Abdominal volume index. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Calculated from anthropometric measurements. Appetite scores. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Visual analogue scale. Systolic blood pressure. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Standard calibrated digital sphygmomanometer after 10 minutes of rest in a seated position. The mean of two measurements will be recorded. Diastolic blood pressure. Timepoint: At baseline and at week 16 after the intervention. Method of measurement: Standard calibrated digital sphygmomanometer after 10 minutes of rest in a seated position. The mean of two measurements will be recorded. National Institute for Medical Research Development (NIMAD) Urmia University of Medical Sciences National Institute for Medical Research Development (NIMAD) Approved 2026-06-16 Ethics Committee of National Institute for Medical Research Development (NIMAD) No. 21, Besat Ave., West Fatemi Ave., Tehran, Iran Tehran Tehran Iran (Islamic Republic of)