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.
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
Allocation
RANDOMIZED
Purpose
TREATMENT
Masking
DOUBLE
Enrollment
140
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.
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.
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.
2 Department of Treatment of Obesity, Metabolic Disorders and Clinical Dietetics, Poznań University of Medical Sciences,
Poznan, Poland
RECRUITINGChanges 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).
Time frame: 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).
Time frame: 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).
Time frame: At the baseline and 12 weeks of treatment
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.
Time frame: 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.
Time frame: 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.
This platform is for informational purposes only and does not constitute medical advice. Always consult a qualified healthcare professional.
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.
Time frame: At the baseline and 12 weeks of treatment
Changes in waist to hip ratio.
WHR will be calculated as WC to HC quotient.
Time frame: 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.
Time frame: 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),
Time frame: 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.
Time frame: 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.
Time frame: At the baseline and 12 weeks of treatment
Gut (taxonomic and functional) microbiota analysis in stool.
Analyzed by the NGS method.
Time frame: 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))
Time frame: 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)
Time frame: At the baseline and 12 weeks of treatment
Changes in ALT, AST, GGT
The ALT, AST, GGT will be measured using standard methods.
Time frame: At the baseline and 12 weeks of treatment
Changes in non-esterified free fatty acids.
The NEFA will be measured using standard methods.
Time frame: 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.
Time frame: 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.
Time frame: At the baseline and 12 weeks of treatment
Changes in fasting glucose level.
The fasting glucose level will be measured using standard methods.
Time frame: At the baseline and 12 weeks of treatment
Changes in fasting insulin level.
The ELISA will be used in the estimation.
Time frame: 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.
Time frame: 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).
Time frame: 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.
Time frame: 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.
Time frame: 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.
Time frame: At the baseline and 12 weeks of treatment
Changes in hsCRP.
The hsCRP will be measured using ELISA.
Time frame: At the baseline and 12 weeks of treatment
Changes in pentraxin 3.
The pentraxin 3 (PTX3) will be measured using ELISA.
Time frame: At the baseline and 12 weeks of treatment
Gut barrier integrity parameter: calprotectin.
The ELISA, will be used in the estimation.
Time frame: 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.
Time frame: At the baseline and 12 weeks of treatment
Gut barrier integrity parameters: lipopolysaccharide (LPS).
The ELISA, will be used in the estimation.
Time frame: 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).
Time frame: At the baseline and 12 weeks of treatment