Effect of Probiotics or Berberine in Hepatic Steatosis Markers, Cardiometabolic and Microbiotic Profile in NAFL.

Effect of Probiotics or Berberine Supplementation on Hepatic Steatosis Markers, Cardiometabolic and Microbiotic Profile in NAFL - A Randomized Double- Blind Clinical Study.

Effect of oral selected Probiotics (PRO) and/or Berberine (BBR) supplementation on hepatic steatosis markers, cardiometabolic profile, and gut microbiota profile in the non-alcoholic fatty liver (NAFL) - a randomized double-blind clinical study.

Study Overview

• Status: Recruiting
• Conditions: Obesity; Non Alcoholic Fatty Liver
• Intervention / Treatment: Dietary supplement: Probiotic; Dietary supplement: Berberine; Dietary supplement: Placebo; Dietary supplement: Probiotc and Berberine

Detailed Description

Probiotics (PRO) and bioactive natural substances such as Berberine (BBR) can improve metabolic parameters in patients with obesity and metabolic disorders. In addition, they significantly affect the composition and function of gut microbiota (GM) and support anti-inflammation and antioxidant defense. These data have become the starting point for the proposed multidirectional approach, aimed at assessing the effect of PRO and/or BBR supplementation on:

• hepatic-related outcomes,
• changes in anthropometric measurements (body mass, BMI, body mass composition and fat mass % content),
• cardiometabolic profile (e.g. blood pressure, noninvasive markers of endothelial function, cardiometabolic biochemical parameters)
• microbiotic profile (gut microbiota composition, endotoxemia)
• the content of the minerals, in overweight/obese patients with nonalcoholic fatty liver (NAFL).

• Study Type: Interventional
• Enrollment (Anticipated): 140
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Małgorzata Moszak, PhD; Phone Number: +48-6185-49-377; Email: mmoszak@ump.edu.pl
• Study Contact Backup: Name: Monika Szulińska, DSc; Phone Number: +48-6185-49-377

Study Locations

• Poland
  • Poznań, Poland, 60-569
    • Recruiting
    • 2 Department of Treatment of Obesity, Metabolic Disorders and Clinical Dietetics, Poznań University of Medical Sciences,
    • Contact: Małgorzata Moszak, PhD; Phone Number: +48-6185-49-377; Email: mmoszak@ump.edu.pl

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 36 years to 56 years (Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

Description

Inclusion Criteria:

• age 40 to 60 years;
• women ≥1 year since last menstruation;
• body mass index (BMI): 27.0 kg/m2 to 34.9 kg/m2;
• abdominal obesity-related waist circumference > 80 cm (women) and >94 cm (men) (in accordance to International Diabetes Federation);
• stable body weight in the 3 months prior to the trial (permissible deviation is ± 3 kg);
• NAFL - diagnosed based on USG in accordance with PGE-NAFLD recommendation

Exclusion Criteria:

• history of following alternative diets within 3 months before the study;
• history of use of any dietary supplements in the 3 months before the study;
• history of intake of antibiotics, probiotics, prebiotics within 3 months before the study;
• secondary form of obesity, pharmacological treatment for obesity (in the 3 months before the study), history of bariatric surgery;
• another liver diseases: high risk of NASH (assessed on the FIB-4, according to the PGE-NAFLD recommendation), autoimmune hepatitis, hepatitis B and C, toxic hepatitis, cirrhosis, Wilson's disease, hemochromatosis;
• other gastrointestinal disorders, especially: IBD, celiac disease, gastritis and duodenitis, pancreatic disorders, gastrointestinal symptoms suggestive of IBS;
• clinically significant acute inflammatory process (elevated hsCRP);
• abnormal kidney function (GFR <60mL/min/1,73m2);
• T2D;
• dyslipidemia or hypertension - requiring the introduction and/or change of pharmacological treatment in the 6 months before the trial or during intervention;
• pump inhibitors, anticoagulants, drugs causing metabolic alteration, e.g., SFAs (second-generation antipsychotics);
• diseases requiring nutritional requirement and chronic supplementation;
• alcohol (>30g/d for men and >20g/d for women), nicotine or drug abuse;
• mental disorders, including eating disorders;
• cancer, autoimmune diseases;
• any other condition which may influence on final results of the study or pose a risk for subjects health.

Study Plan

How is the study designed?

Design Details

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

Number of Arms

4

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Active Comparator: Probiotic; Individuals receive Probiotics (9 strains: B. bifidum W23, B. lactis W51, B. lactis W52, L. acidophilus W37, L. brevis W63, L. casei W56, L. salivarius W24, Lactococcus lactis W19, and Lactococcus lactis W58) in dose: 1x109 colony forming units (CFU), daily, for 12 weeks. Intervention: Dietary Supplement: Probiotic	Dietary supplement: Probiotic; The probiotic group will receive one capsule of the probiotic mixture (dose:1x109 colony forming units (CFU) per day in one dose (before breakfast). The PRO preparation will contain the following bacterial strains: 50% Lactococcus lactis Rosell® - 1058, 25% Lactobacillus casei Rosell® - 215, 12,5% Lactobacillus helveticus Rosell® - 52, 12,5% Bifidobacterium bifidum Rosell® - 71). Probiotics will be administered orally.
Active Comparator: Berberine; Individuals receive Berberine (Berberine hydrochloride 97% extract of Berberis aristata) in dose: 1500 mg/day, for 12 weeks. Intervention: Dietary Supplement: Berberine	Dietary supplement: Berberine; The berberine group will receive 1500 mg of Berberine (Berberine hydrochloride 97% extract of Berberis aristata) per day in 3 doses. Berberine will be administered orally, before breakfast, dinner, and before supper.
Placebo Comparator: Placebo; Individuals receive placebo daily, for 3 months. Intervention: Dietary Supplement: Placebo	Dietary supplement: Placebo; The placebo group will receive a placebo. Placebo will contain only the excipients and will be administered orally for 24 weeks. Placebo in no way: color, taste, smell, form of taking, the dosage will not differ from the preparations tested. However, it will not contain probiotcs or berberine. Placebo will be orally administered three times a day: before breakfast, dinner, and supper (6.00-7.00 p.m.). To meet the GCP conditions, subjects from all groups will receive the same number of capsules (six) per day.
Active Comparator: Probiotics and Berberine; Individuals receive: Probiotics (9 strains: B. bifidum W23, B. lactis W51, B. lactis W52, L. acidophilus W37, L. brevis W63, L. casei W56, L. salivarius W24, Lactococcus lactis W19, and Lactococcus lactis W58) in dose: 1x109 colony forming units (CFU), daily and Berberine (Berberine hydrochloride 97% extract of Berberis aristata) in dose: 1500 mg/day, for 12 weeks. Intervention: Dietary Supplement: Probiotic and Berberine	Dietary supplement: Probiotc and Berberine; Probiotics and Berberine groupwill receive both: a probiotics mixture (as in PRO group: 1x109 CFU/day; in one dose) and 1500 mg/day of Berberine (Berberine hydrochloride 97% extract of Berberis aristata; in 3 doses). Probiotcs and berberine will be administered orally before breakfast, before dinner, and before supper.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Changes in Fibrosis-4 (FIB-4) - Index for Liver Fibrosis.	FIB-4 will be estimated using a medical calculator (based on parameters as: age, ALT, AST, and platelet count).	At the baseline and 12 weeks of treatment
Changes in HSI - Hepatic Steatosis Index.	HSI will be estimated using a medical calculator (based on parameters as: gender, ALT, AST, BMI, and type 2 diabetes).	At the baseline and 12 weeks of treatment
Changes in NAFLD-LFS (liver fat score).	NAFLD-LFS will be estimated using a medical calculator (based on serum aspartate transaminase/alanine transaminase (AST/ALT) ratio, fasting serum aspartate transaminase (AST) level, fasting serum insulin level, presence of metabolic syndrome and diabetes mellitus).	At the baseline and 12 weeks of treatment

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Changes in blood pressure.	Resting seated BP (both systolic and diastolic) will be measured using a brachial cuff (Omron Healthcare, Kyoto, Japan) three times at 2-min intervals, and the average value will be calculated.	At the baseline and 12 weeks of treatment
Changes in weight.	Weight will be measured in light clothing and without metal objects (i.e., belt, jewelry). Weight will be measured to the nearest 0.1 kg.	At the baseline and 12 weeks of treatment
Changes in waist circumference, hip circumference.	WC will be measured between the iliac crest and the lower rib at the end of a normal expiration, and HC will be measured around the widest portion of the buttocks. Both HC and WC will be measured using stretch-resistant medical tape (Seco) to the nearest 0.5 cm. The measurement of WC and HC will be performed according to the World Health Organization (WHO) protocol.	At the baseline and 12 weeks of treatment
Changes in waist to hip ratio.	WHR will be calculated as WC to HC quotient.	At the baseline and 12 weeks of treatment
Changes in BMI.	BMI will be calculated according to the formula: BMI = kg/m2 where kg is a person's weight in kilograms and m2 is their height in meters squared.	At the baseline and 12 weeks of treatment
Changes in fat mass content in the body.	The fat mass content [in kg] in the body will be estimated using BIA methods (InBody 270 and Bioscan 920-2),	At the baseline and 12 weeks of treatment
Changes in pulse wave velocity (PWV).	The pulse wave velocity (PWV) will be measured using the SphygmoCor Px (Atcor Medical Blood Pressure Analysis System, Sydney, Australia) in a temperature-controlled room before making anthropometric measurements and blood collection.	At the baseline and 12 weeks of treatment
Changes in pulse wave analysis (PWA).	The pulse wave analysis (PWA)will be measured using the SphygmoCor Px (Atcor Medical Blood Pressure Analysis System, Sydney, Australia) in a temperature-controlled room before making anthropometric measurements and blood collection.	At the baseline and 12 weeks of treatment
Gut (taxonomic and functional) microbiota analysis in stool.	Analyzed by the NGS method.	At the baseline and 12 weeks of treatment
Short-chain fatty acids (SCFAs) concentration in stool.	Analyzed using gas chromatography (Agilent Technologies 1260 A GC system with a flame ionization detector (FID))	At the baseline and 12 weeks of treatment
Measurement of hair minerals (Fe, Mg, Ca, Cu, Zn) concentration.	Performed using atomic absorption spectrometry (Atomic Absorption Spectrophotometer ZA3000, Hitachi, Tokyo, Japan)	At the baseline and 12 weeks of treatment
Changes in ALT, AST, GGT	The ALT, AST, GGT will be measured using standard methods.	At the baseline and 12 weeks of treatment
Changes in non-esterified free fatty acids.	The NEFA will be measured using standard methods.	At the baseline and 12 weeks of treatment
Changes in lipids profile (TC, HDL, TG).	The TC, HDL, TG will be measured using standard methods.	At the baseline and 12 weeks of treatment
Changes in low-density lipoprotein (LDL).	The low-density lipoprotein cholesterol (LDL) will be calculated according to the Friedwald equation.	At the baseline and 12 weeks of treatment
Changes in fasting glucose level.	The fasting glucose level will be measured using standard methods.	At the baseline and 12 weeks of treatment
Changes in fasting insulin level.	The ELISA will be used in the estimation.	At the baseline and 12 weeks of treatment
Changes in insulin resistance index (HOMA-IR)	The HOMA-IR will be calculated according to formula: HOMA-IR = (insulin * glucose) / 22.5 for the glucose concentration in mmol/L, or: HOMA-IR = (insulin * glucose ) / 405 for glycemia in mg/dL. In both cases, the insulin is in mU/L.	At the baseline and 12 weeks of treatment
Changes in parameter of liver damage: cytokeratin 18.	The ELISA will be used in the estimation cytokeratin 18 (ccK18).	At the baseline and 12 weeks of treatment
Changes in parameter of liver damage: Glutathione S-transferase (GST).	The ELISA will be used in the estimation GST.	At the baseline and 12 weeks of treatment
Changes in parameter of liver damage: collagen IV.	The ELISA will be used in the estimation collagen IV.	At the baseline and 12 weeks of treatment
Changes in parameter of liver damage: hyaluronic acid.	The ELISA will be used in the estimation hyaluronic acid.	At the baseline and 12 weeks of treatment
Changes in hsCRP.	The hsCRP will be measured using ELISA.	At the baseline and 12 weeks of treatment
Changes in pentraxin 3.	The pentraxin 3 (PTX3) will be measured using ELISA.	At the baseline and 12 weeks of treatment
Gut barrier integrity parameter: calprotectin.	The ELISA, will be used in the estimation.	At the baseline and 12 weeks of treatment
Gut barrier integrity parameters: liver fatty acid-binding protein (L-FABP), intestinal fatty acid-binding protein (I-FABP).	The ELISA, will be used in the estimation.	At the baseline and 12 weeks of treatment
Gut barrier integrity parameters: lipopolysaccharide (LPS).	The ELISA, will be used in the estimation.	At the baseline and 12 weeks of treatment
Cardiometabolic risk.	Cardiometabolic risk will be estimated at baseline and after 3 months of treatment using the SCORE scale. SCORE scale summarize 5 risk factors (sex, age, systolic blood pressure, total cholesterol and smoking).	At the baseline and 12 weeks of treatment

Collaborators and Investigators

• Sponsor: Poznan University of Medical Sciences
• Investigators: Principal Investigator: Paweł Bogdański, Professor, Poznan University of Medical Sciences, Poznan, Poland

Study record dates

Study Major Dates

• Study Start (Actual): August 2, 2022
• Primary Completion (Anticipated): December 31, 2023
• Study Completion (Anticipated): June 1, 2024

Study Registration Dates

• First Submitted: April 25, 2022
• First Submitted That Met QC Criteria: August 29, 2022
• First Posted (Actual): August 31, 2022

Study Record Updates

• Last Update Posted (Actual): August 31, 2022
• Last Update Submitted That Met QC Criteria: August 29, 2022
• Last Verified: August 1, 2022

More Information

Terms related to this study

• Keywords: NAFL; obesity; probiotics; berberine; microbiota
• Additional Relevant MeSH Terms: Digestive System Diseases; Liver Diseases; Fatty Liver; Non-alcoholic Fatty Liver Disease
• Other Study ID Numbers: 491/20

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