Probiotic Intervention, Bile Acids and Lipid Metabolism Metabolic Profile in Healthy Overweight Subjects. (PROBILIP)

Effects of the Probiotic AB-LIFE on the Amount and Profile of Bile Acids, and the Lipid and Metabolic Profile in Healthy Overweight Subjects.

Moderate hypercholesterolemia, metabolic alterations derived from overweight or obesity, and consequently and their related cardiovascular risks can be reduced through changes in lifestyle. A growing body of evidence shows a relationship between hypercholesterolemia and dysfunction of the gut microbiota.

Gut microbiota is considered a keystone in maintaining the health condition of the host through multiple mechanisms affecting different metabolic processes, including lipid metabolism and cholesterol-related pathways. Bile acids (BA) are cholesterol-derived compounds synthesized in the liver and metabolized upon modification by gut bacteria once they reach the colon.

Conversely, BAs shape the composition and function of the intestinal microbiota. This mutual interplay between BAs and gut microbiota regulates many physiological processes, including the lipid, carbohydrate and energy metabolism of the host.

Previous studies based on Lactobacillus plantarum have shown 3 strains CECT 7527, 7528, and 7529 with the ability to adhere to the intestinal mucosa with very low toxicity, what makes them susceptible to be used as a probiotic. The lipid-lowering effect of the 3 bacteria strains was already evidenced in previous preclinical studies in animal models and clinical studies in hypercholesterolemic subjects. Up to now, however, little is known about the effects of the 3- combined Lactobacillus plantarum strains on levels and profile of bile acids in healthy overweight subjects, otherwise at low cardiovascular risk.

The aim of this study is to explore on the mechanism of action of a Lactobacillus plantarum mixture (CECT 7527, CECT 7528 and CECT 7529) and evaluate the effect of this probiotic formulation on BA profile as well as on plasma lipids and other related biomarkers when administered in a dose-dependent regime in a cohort of overweight subjects.

The probiotic product was administrated for 4 weeks with a weekly dose-regime of 2x, 3x, and 4x, respectively, in the second, third, and fourth week in relation to the first week. The effects were evaluated on: (1) level and profile of bile acids in plasma and feces, (2) plasma lipid/lipoprotein and fatty acid profile and (3) endocrine hormones, glucose metabolism and inflammatory markers in plasma.

Study Overview

• Status: Completed
• Conditions: Probiotic Intervention
• Intervention / Treatment: Dietary supplement: Probiotic preparation of three Lactobacillus plantarum strains (CECT 7527, CECT 7528, CECT 7529)

Detailed Description

Sample size (N=20) was calculated according results of previous studies on plasma bile acids. The study refers to healthy overweight [body mass index (BMI) 25.0-29.9 kg/m2] adult men (N=10) and women (N=10) between the ages of 25 and 60 years.

The study was approved by the Human Ethical Review Committee of the Hospital Sant Pau in Barcelona (register number: 20/029). Informed written consent was obtained from all participants before their inclusion in the study. To confirm health status, all subjects underwent a complete physical examination conducted by the study physician.

The study lasted 6 weeks that were structured in:

• 2 weeks of run-in.

• 4 weeks of intervention period divided into 4 phases of 7 days. During the intervention period, Lactobacillus plantarum capsules were consumed according to the following pattern:

  • Days 0-6: 1 capsule/day
  • Days 7-13: 2 capsules/day
  • Days 14-20: 3 capsules/day
  • Days 21-27: 4 capsules/day

The volunteers visited the center at the beginning of each phase of the intervention period (Days 0, 7, 14 and 21) and at the end of the intervention period (Day 28).

Dietary habits were collected. Compliance was monitored by weekly telephone contact with participants and interviewing them at the end of each intervention period. Blood samples were collected early at the morning after twelve-hour fasting at baseline and at the end of each intervention phase. Blood samples were used for determining all variables of the study. Stool samples were obtained at baseline and after 28 days intervention for the study of bile acids, fatty acids and microbiota.

• Study Type: Interventional
• Enrollment (Actual): 20
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Spain
  • Barcelona, Spain, 08025
    • Institut Recerca-Hospital Santa Creu I Sant Pau

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 25 years to 60 years (Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

Description

Inclusion Criteria:

• Body Mass index (BMI): Between 25 and 30 Kg / m2

Exclusion Criteria:

• Eating disorders.
• Subjects with cardiovascular risk factors such as hypercholesterolemia (genetic or secondary), hypertension and diabetes, treated pharmacologically.
• History of ischemic heart disease (and / or previous angina or AMI) or arrhythmia (current or previous).
• Previous strokes and / or peripheral vascular disease.
• Alcohol consumption greater than 60 gr / day.
• Kidney failure (creatinine> 2 mg / dl).
• Presence of neoplasia.
• Presence of systemic disease.
• Psychiatric illness under treatment with psychotropics.
• Unstabilized thyroid disease.
• Having followed or being following a hypocaloric diet or regularly consumed dietary supplements for weight or constipation or cholesterol control (plant sterols, soy lecithin, omega-3 fatty acids) for 2 months prior to inclusion in the study.
• Pregnancy or breastfeeding.
• In current treatment with non-steroidal anti-inflammatory drugs, antiplatelet agents, fibrates or statins.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: N/A
• Interventional Model: Single Group Assignment
• Masking: None (Open Label)

Number of Arms

1

Arms and Interventions

Participant Group / Arm

Intervention / Treatment

Experimental: Probiotic mixture; Arm that received Lactobacillus plantarum mixture (CECT 7527, CECT 7528 and CECT 7529)

Dietary supplement: Probiotic preparation of three Lactobacillus plantarum strains (CECT 7527, CECT 7528, CECT 7529); All subjects were submitted to 4 weeks intervention consisting on a dose-dependent regime of Lactobacillus plantarum AB-B (strains CECT 7527, CECT 7528, CECT 7529; AB-LIFE). Before starting the intervention period, all subjects were submitted to a 2-weeks run-in period. During the wash-in and intervention-periods, participants were instructed to maintain their habitual dietary habits and to continue their normal pattern of physical activity throughout the study period. Treatments during the intervention period were: Week 1: 1 capsule/day taken during breakfast; Week 2: 2 capsules / day at- breakfast; Week 3: 2 capsules at breakfast and 1 at dinner; Week 4: 2capsules at breakfast and 2 capsules at dinner. Each capsule contains 1.2x10^9 cfu (colony forming units) in a 1: 1: 1 ratio of the three strains.; Other Names: AB-LIFE

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Change from Baseline Lipid profile at 7 days	By standardized biochemical techniques	At days 0 and 7
Change from Baseline Lipid profile at 14 days	By standardized biochemical techniques	At days 0 and 14
Change from Baseline Lipid profile at 21 days	By standardized biochemical techniques	At days 0 and 21
Change from Baseline Lipid profile at 28 days	By standardized biochemical techniques	At days 0 and 28
Change from Baseline Serum bile acids at 14 days	By ultra-high performance liquid chromatography - mass spectrometry (UHPLC-MS)	At days 0 and 14
Change from Baseline Serum bile acids at 28 days	By ultra-high performance liquid chromatography - mass spectrometry (UHPLC-MS)	At days 0 and 28
Change from Faecal bile acids at 28 days	By ultra-high performance liquid chromatography - mass spectrometry (UHPLC-MS)	At days 0 and 28
Change from Baseline Serum fatty acids at 14 days	By ultra-high performance liquid chromatography - mass spectrometry (UHPLC-MS)	At days 0 and 14
Change from Baseline Serum fatty acids at 28 days	By ultra-high performance liquid chromatography - mass spectrometry (UHPLC-MS)	At days 0 and 28
Change from Baseline Faecal fatty acids at 28 days	By ultra-high performance liquid chromatography - mass spectrometry (UHPLC-MS)	At days 0 and 28
Change from Baseline Serum lipoprotein profile at 14 days	By nuclear magnetic resonance (NMR)	At days 0 and 14
Change from Baseline Serum lipoprotein profile at 28 days	By nuclear magnetic resonance (NMR)	At days 0 and 28
Change from Baseline Plasmatic endocrine hormones and metabolic markers at 14 days	By Enzyme-Linked immunosorbent Assay (ELISAs)	At days 0 and 14
Change from Baseline Plasmatic endocrine hormones and metabolic markers at 28 days	By Enzyme-Linked immunosorbent Assay (ELISAs)	At days 0 and 28

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Change from Baseline Plasmatic Inflammatory markers at 14 days	By standardized biochemical techniques and Multiplex for Luminex Immunoassays	At days 0 and 14
Change from Baseline Plasmatic Inflammatory markers at 28 days	By standardized biochemical techniques and Multiplex for Luminex Immunoassays	At days 0 and 28
Change from Baseline LDL susceptibility to oxidation and HDL antioxidant capacity at 14 days	By conjugated dienes measurement and by Total radical-trapping antioxidant potential (TRAP) respectively	At days 0 and 14
Change from Baseline LDL susceptibility to oxidation and HDL antioxidant capacity at 28 days	By conjugated dienes measurement and by Total radical-trapping antioxidant potential (TRAP) respectively	At days 0 and 28
Change from Baseline Serum Vitamin D and thyroid hormones (TSH, T3 & T4) at 14 days	By Liquid Chromatography with tandem mass spectrometry (LC-MS/MS) and by chemoluminescence immunoassays respectively	At days 0 and 14
Change from Baseline Serum Vitamin D and thyroid hormones (TSH, T3 & T4) at 28 days	By Liquid Chromatography with tandem mass spectrometry (LC-MS/MS) and by chemoluminescence immunoassays respectively	At days 0 and 28

Collaborators and Investigators

• Sponsor: Fundació Institut de Recerca de l'Hospital de la Santa Creu i Sant Pau
• Collaborators: AB Biotics, SA

Study record dates

Study Major Dates

• Study Start (Actual): September 1, 2020
• Primary Completion (Actual): November 28, 2020
• Study Completion (Actual): February 21, 2022

Study Registration Dates

• First Submitted: May 5, 2022
• First Submitted That Met QC Criteria: May 12, 2022
• First Posted (Actual): May 18, 2022

Study Record Updates

• Last Update Posted (Actual): June 7, 2022
• Last Update Submitted That Met QC Criteria: June 3, 2022
• Last Verified: June 1, 2022

More Information

Terms related to this study

• Keywords: Inflammation; Microbiota; Lipids; Metabolism; Bile acids
• Additional Relevant MeSH Terms: Body Weight; Overweight
• Other Study ID Numbers: IIBSP-PRO-2019-122

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No