Red Wine Effects Upon Gut Flora and Plasma Levels of Trimethylamine-N-oxide (TMAO) - WineFlora Study

Sponsor

University of Sao Paulo General Hospital (Other)

Overall Status

Completed

CT.gov ID

NCT03232099

Collaborator

Fundação de Amparo à Pesquisa do Estado de São Paulo (Other), IBRAVIN (Other)

Enrollment

Arms

19.4

Actual Duration (Months)

Study Details

Study Description

Brief Summary

Recent evidence indicates that Trimethylamine-N-oxide (TMAO) is a pro-atherosclerotic, phosphatidylcholine-dependent metabolite of diet and intestinal flora. Food substrates derive from carnitine and phosphatidylcholine (lecithin), present mainly in eggs, red meat, liver and pork. The intestinal flora pattern that favors the formation of TMAO is very similar to that which predisposes to insulin resistance and obesity: a high proportion between phylum Firmicutes over Bacteroidetes. The intestinal microbiota is sensitive and variable; the use of prebiotics and probiotics can change the relationship between Firmicutes/Bacteroidetes phyla. Red wine (RW), for its composition with polyphenols and possible bactericidal role, may play a role in the intestinal flora modification and could promote proliferation of beneficial bacteria. However, the influence of RW on TMAO is not known. This is the hypothesis to be tested in this trial. METHODS: This is a prospective, crossover, randomized, controlled trial with patients from Heart Institute (InCor), FMUSP and volunteers recruited through press releases. We will evaluate 42 patients, all men, with established atherosclerotic disease. Patients will be evaluated in a crossed manner: each subject receives both treatments, intervention and control (in random order), and they will be divided into 2 groups: A and B. In the first intervention stage, after 2 weeks of washout for all patients , group A receives Red Wine (RW) and group B is the control, abstemious. In the 2nd stage of intervention, after 2 weeks of washout for all patients the groups are inverted: group B receives RW; and group A will be abstemious. In the period with wine intervention, patients will receive 250 mL/day of red wine per day, for 5 days of the week, for 3 weeks. Patients will maintain their usual diet without the use of prebiotics or probiotics, or other polyphenolic derivatives. At the beginning and at the end of each stage, patients will be submitted to serum TMAO and intestinal microbiota evaluation. For the intestinal microbiota evaluation, the new generation sequencing will be used in the highly preserved portion of the 16S subunit of the rRNA gene. The determination of TMAO in plasma will be by liquid chromatography coupled to mass spectrometry. Expected results: It is expected to determine if RW acts on the intestinal flora to the point of influencing plasma TMAO concentration.

Condition or Disease	Intervention/Treatment	Phase
Trimethylamine-N-oxide Gut Microbiota Effects of Red Wine Atherosclerosis	Dietary Supplement: Red Wine consumption Other: Period without alcohol consumption	N/A

Study Design

Study Type:

Interventional

Actual Enrollment :

42 participants

Allocation:

Randomized

Intervention Model:

Crossover Assignment

Intervention Model Description:

Forty two men with established atherosclerotic disease will be studied:after a 2 week washout period, each subject receives both treatments, intervention and controlForty two men with established atherosclerotic disease will be studied:after a 2 week washout period, each subject receives both treatments, intervention and control

Masking:

None (Open Label)

Primary Purpose:

Basic Science

Official Title:

Evaluation of Red Wine Effects Upon Gut Flora and Plasma Levels of Trimethylamine-N-oxide (TMAO) in Patients With Established Atherosclerotic Disease

Actual Study Start Date :

Aug 31, 2016

Actual Primary Completion Date :

Mar 22, 2018

Actual Study Completion Date :

Apr 15, 2018

Arms and Interventions

Arm	Intervention/Treatment
Experimental: Red Wine group (RW) After a two weeks washout period,in the intervening period with red wine, patients will receive 250 mL / day of red wine per Day, 5 days a week, for 3 weeks.	Dietary Supplement: Red Wine consumption After a two weeks washout period,in the intervening period with red wine, patients will receive 250 mL / day of red wine per Day, 5 days a week, for 3 weeks.
Active Comparator: Abstemious After a two weeks washout period,in the Abstemious period, patients should not consume any kind of alcohol beverages, for 3 weeks	Other: Period without alcohol consumption After a two weeks washout period,in the Abstemious period, patients should not consume any kind of alcohol beverages, for 3 weeks Other Names: Abstain Period

Arm

Intervention/Treatment

Experimental: Red Wine group (RW)

After a two weeks washout period,in the intervening period with red wine, patients will receive 250 mL / day of red wine per Day, 5 days a week, for 3 weeks.

Dietary Supplement: Red Wine consumption

After a two weeks washout period,in the intervening period with red wine, patients will receive 250 mL / day of red wine per Day, 5 days a week, for 3 weeks.

Active Comparator: Abstemious

After a two weeks washout period,in the Abstemious period, patients should not consume any kind of alcohol beverages, for 3 weeks

Other: Period without alcohol consumption

After a two weeks washout period,in the Abstemious period, patients should not consume any kind of alcohol beverages, for 3 weeks

Other Names:

Abstain Period

Outcome Measures

Primary Outcome Measures

Gut flora modification assessed by 16S ribosomal rNA gene sequencing and plasma concentration of TMAO changes measured by liquid chromatography coupled to mass spectrometry in 42 patients after ingestion 250 mL of Red Wine 5 days/week, for 3 weeks [up to 10 weeks]
Patients will be divided into 2 groups: A and B. In the first intervention stage, after 2 weeks of washout for all patients , group A receives Red Wine (RW) and group B is the control, abstemious. In the 2nd stage of intervention, after 2 weeks of washout for all patients the groups are inverted: group B receives RW; and group A will be abstemious. In the period with wine intervention, patients will receive 250 mL/day of red wine per day, for 5 days of the week, for 3 weeks. Patients will maintain their usual diet without the use of prebiotics or probiotics, or other polyphenolic derivatives. At the beginning and at the end of each stage, patients will be submitted to serum TMAO and intestinal microbiota evaluation. For the intestinal microbiota evaluation, the new generation sequencing will be used in the highly preserved portion of the 16S subunit of the rRNA gene. The determination of TMAO in plasma will be by liquid chromatography coupled to mass spectrometry

Eligibility Criteria

Criteria

Ages Eligible for Study:

45 Years to 70 Years

Sexes Eligible for Study:

Male

Accepts Healthy Volunteers:

Inclusion Criteria:

Men, 45-70 years
Established atherosclerotic diseases defined as:

Previous history of CAD: angina or acute myocardial infarction (AMI), myocardial revascularization or angioplasty, angiographic evidence of stenosis ≥50% in at least one of the major coronary arteries B. Cerebrovascular disease: transient ischemic stroke or cerebrovascular accident C. Peripheral arterial disease: clinical evidence of extracoronary atherosclerosis

Exclusion Criteria:

Acute events in the last 30 days (AMI, troponin elevation, coronary angioplasty or coronary artery bypass grafting)
Heart Failure (NYHA functional class ≥ II)
Renal Failure (clearance creatinine <30 mL / min by the Cockcroft-Gault formula)
Hepatic Failure
Gastro-intestinal cancer
Intestinal inflammatory diseases
Obstructive biliary diseases
Prior gastrointestinal surgeries: cholecystectomy or colectomy
Use of antibiotics within the last 2 months or during protocol
Alcoholism or alcohol intolerance
Diabetes mellitus or use of antidiabetics drugs

Contacts and Locations

Locations

No locations specified.

Sponsors and Collaborators

University of Sao Paulo General Hospital
Fundação de Amparo à Pesquisa do Estado de São Paulo
IBRAVIN

Investigators

Study Chair: Protasio L da Luz, Professor, Instituto do Coração - InCor - University of Sao Paulo General Hospital
Principal Investigator: Elisa A Haas, MD, Instituto do Coração - InCor - University of Sao Paulo General Hospital