The Effects of Fat and Theobromine on apoA-I

The Effects of a Diet Rich in Fat or Theobromine on Intestinal apoA-I mRNA and Protein Expression

Rationale: Despite successful efforts to lower atherogenic serum low-density lipoprotein (LDL) cholesterol concentrations, a substantial residual cardiovascular risk is still present. An additive strategy to further lower this residual risk may be via raising high-density lipoprotein (HDL) cholesterol concentrations, and in particular those of its major protein constituent apolipoprotein A-I (apoA-I). However, recent evidence shows that raising HDL cholesterol (HDL-C) concentrations not always causes a reduction in cardiovascular disease (CVD) risk. To understand this it is important to know what are the targets and molecular mechanisms that underlie a shift to a HDL fraction with cardioprotective activities. There is increasing evidence that particularly the apoA-I proteins in the HDL fractions are atheroprotective. It is important to verify whether the effect of an intervention actually increases apoA-I production since that results in the formation of de-novo small pre-beta HDL particles that have full capacity to resorb cholesterol from the arterial wall and return this to the liver for secretion (reverse cholesterol transport). The investigators here propose to evaluate, in a double-blind human intervention study, the difference in intestinal apoA-I gene expression, of healthy subjects, after consumption of a high fat (HF) or low fat (LF) meal. Additionally the investigators propose to evaluate the effect of theobromine on intestinal apoA-I expression. Based on several studies, theobromine improved cardiovascular risk parameters among which an elevation in apoA-I and HDL cholesterol concentrations. It is however unknown whether this effect of theobromine originates from elevated apoA-I production, which suggest improved functionality, or decreased clearance, suggesting reduced functionality. The investigators therefore propose to also evaluate if the usage of theobromine is a better way to increase intestinal apoA-I mRNA and protein expression, than consumption of a HF meal. The investigators will do this in the same double-blind human intervention study. The theobromine will be added in a third arm in a LF condition.

Study design: The proposed study will have a randomized, double-blind cross-over design. The total study duration will be 17 days, consisting of 3 experimental days with postprandial tests, each separated by a 1 week wash-out period.

Study population: Ten apparently healthy male subjects, aged 18-60 years, without a history of any gastrointestinal disorders or complaints.

Intervention: During the three experimental days, subjects will consume a milkshake: one HF milkshake, one LF milkshake and one LF milkshake supplemented with 850 mg theobromine. Total follow-up during each of the postprandial tests is 5 hours.

Study Overview

• Status: Completed
• Conditions: Cardiovascular Diseases
• Intervention / Treatment: Dietary supplement: Fat milkshake; Dietary supplement: Theobromine

• Study Type: Interventional
• Enrollment (Actual): 10
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Netherlands
  • Maastricht, Netherlands, 6202
    • Maastricht University Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 60 years (Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: Male

Description

Inclusion Criteria:

• Aged between 18 and 60 years
• Serum total cholesterol <8.0 mmol/L
• Plasma glucose <7.0 mmol/L
• BMI between 20 - 30 kg/m2
• Non-smoking

• Willingness to abstain from theobromine rich products from two weeks prior to the study maintaining throughout the full duration (17 days) of the study:

  • Chocolate containing products (see Appendix A).
• Willingness to abstain from energy drinks from two weeks prior to the study maintaining throughout the full duration (17 days) of the study, because of its high caffeine content,

• Willingness to reduce caffeine containing drinks till maximum 4 drinks a day from two weeks prior to the study maintaining throughout the full duration (17 days) of the study:

  • Coffee
  • Tea
  • Cola.

Exclusion Criteria:

• Unstable body weight (weight gain or loss >3 kg in the past 3 months),
• Not willing to stop the consumption of gastric acid inhibitor, laxantia, prebiotics, probiotics and antibiotics for at least one month before the start of the study and during the study.
• Any medical condition requiring treatment and/or medication use,
• Indication for treatment with cholesterol-lowering drugs according to the Dutch Cholesterol Consensus (35)
• Use of medication or a medically-prescribed diet known to affect serum lipid or glucose metabolism. The use of oral contraceptives and paracetamol is allowed.
• Active cardiovascular disease (for instance congestive heart failure) or recent (<6 months) event, such as acute myocardial infarction or cerebro-vascular accident.
• Gastrointestinal diseases (like celiac disease, inflammatory bowel disease, irritable bowel disease and food allergies) or a history of any gastrointestinal disorders or complaints.
• Severe medical conditions that might interfere with the study such as epilepsy, asthma, chronic obstructive pulmonary disease and rheumatoid arthritis.
• Not willing to stop the consumption of vitamin supplements or fish oil capsules 3 weeks before the start of the study
• Consumption of >21 glasses of alcohol-containing drinks per week.
• Abuse of drugs
• Participation in another biomedical study within 1 month prior to the screening visit
• Having donated > 150 ml blood within 1 month prior to the screening visit, planning to donate blood during the study or within one month after finishing the study.
• Impossible or difficult to puncture as evidenced during the screening visits.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Prevention
• Allocation: Randomized
• Interventional Model: Crossover Assignment
• Masking: Triple

Number of Arms

3

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: High fat meal; A high fat milkshake containing 60.6 grams of fat	Dietary supplement: Fat milkshake
Placebo Comparator: Low fat meal; A Low fat milkshake containing 10.5 gram of fat	Dietary supplement: Fat milkshake
Experimental: A Low fat meal containing theobromine; A low fat milkshake containing 10.5 grams of fat supplemented with 850 mg of theobromine	Dietary supplement: Fat milkshake; Dietary supplement: Theobromine

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Microarray	On each 3 test days 5 hours after the postprandial meal a biopsy will be taken. From these intestinal cells gene expression profiles will be measured by microarray.	up to 3 days

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Blood samples	During each test day blood samples will be taken in intervals after consumption of the postprandial meal. Blood samples are taken to analyse lipids, apoA-I, apoB-100, apoB48, CRP, cholesterol efflux, glucose and free fatty acids.	Day 1, 2 and 3 at T=0, 15, 30, 45, 60, 90, 120, 180 and 240 min

Collaborators and Investigators

• Sponsor: Maastricht University Medical Center
• Investigators: Principal Investigator: Jogchum Plat, Prof, Maastricht University Hospital; Study Director: Ronald Mensink, Prof, Maastricht University Hospital

Study record dates

Study Major Dates

• Study Start: February 1, 2014
• Primary Completion (Actual): July 1, 2014
• Study Completion (Actual): July 1, 2014

Study Registration Dates

• First Submitted: February 17, 2014
• First Submitted That Met QC Criteria: March 11, 2014
• First Posted (Estimate): March 12, 2014

Study Record Updates

• Last Update Posted (Estimate): August 5, 2014
• Last Update Submitted That Met QC Criteria: August 4, 2014
• Last Verified: August 1, 2014

More Information

Terms related to this study

• Keywords: Cardiovascular Diseases; HDL metabolism; ApoA-I; High fat; Theobromine
• Additional Relevant MeSH Terms: Cardiovascular Diseases; Physiological Effects of Drugs; Vasodilator Agents; Autonomic Agents; Peripheral Nervous System Agents; Bronchodilator Agents; Anti-Asthmatic Agents; Respiratory System Agents; Theobromine
• Other Study ID Numbers: METC 13-3-049