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Differential Effects of Eicosapentaenoic Acid (EPA) and Docosahexaenoic Acid (DHA) on Platelet, Endothelial and Vascular Function

Not Applicable
Completed
Conditions
Healthy
Interventions
Dietary Supplement: Placebo - olive oil (BP specification)
Dietary Supplement: DHA-rich triacylglycerol oil
Dietary Supplement: EPA-rich triacylglycerol oil
Registration Number
NCT01735357
Lead Sponsor
King's College London
Brief Summary

The purpose of this study was to determine whether supplementation with oils enriched with long chain n-3 PUFA, either EPA or DHA, had a differential effect on platelet, endothelial and vascular function.

Detailed Description

Relatively few studies have made a head-to-head comparison of DHA (22:6n-3) with EPA (20:5n-3). The understanding of this differential effect may be of great interest in populations with low EPA intake such as vegetarians, who may choose to supplement their dietary intake of long-chain n-3 PUFA in the form of DHA-rich algal oil.

This study aimed to investigate the effect of supplementation with oils rich in either EPA or DHA (3g/day, 6 weeks) in healthy young males on platelet, endothelial and vascular function, as well as other CVD risk factors. The primary outcomes were platelet monocyte aggregates and endothelial progenitor cells - novel markers of platelet and endothelial function, measured by flow cytometry, Secondary outcomes included capillary density, measured by capillaroscopy to assess changes in microvascular function, pulse wave analysis, digital volume pulse and ambulatory blood pressure. Other secondary outcomes included lipid profiles (TAG, cholesterol, NEFA), glycaemic control (HOMA, QUICKI) and oxidative stress (isoprostane). The omega-3 index (erythrocyte EPA+DHA) was used as a marker of compliance.

Recruitment & Eligibility

Status
COMPLETED
Sex
Male
Target Recruitment
48
Inclusion Criteria
  • Healthy males
  • No smokers
  • Aged 18-45y old
  • Able to understand the information sheet and comply with all the trial procedures
  • Having given written consent to take part in the study prior to participation.
Exclusion Criteria
  • Reported history of CVD (myocardial infarction, angina, venous thrombosis, stroke, dyslipidemia), diabetes (or fasting glucose ≥ 6.1 mmol/L), cancer, kidney, liver or bowel disease.
  • Presence of gastrointestinal disorder or use of drug, which is likely to alter gastrointestinal motility or nutrient absorption.
  • Current smokers; history of substance abuse or alcoholism (previous weekly alcohol intake >60 units/week); current self-reported weekly alcohol intake exceeding 28 units
  • Recent use of hypolipidaemic, antihypertensive, antiplatelet or antithrombotic mediations
  • Platelet count above or below the normal range or any history indicative of a congenital or acquired platelet or haemostatic defect.
  • Allergy or intolerance to any component of study capsules
  • Unwilling to restrict consumption of any source of fish oil for the length of the study
  • Subjects reporting consumption of >1 portion oily fish per week
  • Weight change of >3 kg in preceding 2 months; BMI <18 and >32 kg/m2
  • Blood pressure>160/90 mmHg
  • Fasting blood cholesterol > 6.5 mmol/L; fasting triacylglycerol concentrations > 2.0 mmol/L

Study & Design

Study Type
INTERVENTIONAL
Study Design
PARALLEL
Arm && Interventions
GroupInterventionDescription
Olive oil (BP specification)Placebo - olive oil (BP specification)5g per day
DHA-rich oilDHA-rich triacylglycerol oilFish oil supplement (total = 5g/day) providing 3.1g/day of DHA triacylglycerol, blended with olive oil
EPA-rich oilEPA-rich triacylglycerol oilFish oil supplement (total = 5g/day) providing 2.9g/day of EPA triacylglycerol, blended with olive oil
Primary Outcome Measures
NameTimeMethod
Platelet Monocyte Aggregates (PMA)6 weeks

The endothelium plays a vital role in the regulation of blood flow, thrombosis and inflammation. Endothelium-derived anti-adhesive and anti-aggregant substances, including prostacyclin and nitric oxide, are known to inhibit platelet activation. Endothelial dysfunction or vessel wall injury lead to the activation of platelets, of which platelet-monocyte-aggregates (PMA) are a sensitive marker, and were shown to inversely correlate with markers of EF in patients with stable CHD. The measurement of PMA by flow cytometry is a method which reduces ex vivo platelet activation to its minimum and is believed to represent platelet activation in vivo.

Endothelial Progenitor Cell (EPC) counts6 weeks

EPCs are a subgroup of circulating progenitor cells that are recruited from the bone marrow to repair the injured vasculature. They have been associated with a reduced CVD risk and may serve as markers of endothelial function because they represent a greater capacity for the endothelium to repair itself. Two populations of EPCs were measured by flow cytometry, described as 'early EPC' (KDR+/CD34+/CD133+) and 'late EPCs' (KDR+/CD34+/CD31+).

Secondary Outcome Measures
NameTimeMethod
Serum total cholesterol concentration6 weeks
Serum high density lipoprotein concentration6 weeks
Arterial stiffness6 weeks

Pulse wave analysis (PWA) was used to measure indices of arterial stiffness, including peripheral and central augmentation index, as well as central systolic and diastolic blood pressure. Digital volume pulse (DVP) was used to measure reflection and stiffness indices.

Blood Pressure (BP) and Heart Rate (HR)6 weeks

Resting HR and BP were measured in the seated and supine position after 15 minute rest, on the days of both baseline and endpoint visits. In addition, ambulatory BP and HR (24h, daytime, nighttime) was measured 2-3 days prior to each visit.

Plasma Nitrate and Nitrites (NOx) concentrations6 weeks

Plasma NOx was measured as a circulating marker of endothelial function.

Serum Triacylglycerol concentrations6 weeks
Serum non esterified fatty acids (NEFA) concentration6 weeks
Serum Apolipoprotein B concentration6 weeks
Plasma glucose concentration6 weeks
Serum resistin concentration6 weeks
Fatty acid profile of plasma non esterified fatty acid fraction6 weeks
Capillary density6 weeks

Capillary rarefaction has been associated to CVD risk factors such as hypertension, smoking and obesity. The cutaneous circulation has emerged as an accessible and representative vascular bed to look at microvascular dysfunction. Capillary density was measured by a Capiscope (kk technologies)

Plasma isoprostane concentrations6 weeks

8-iso-prostaglandin-F2α (8-IsoP-F2α), a prostaglandin F2-like compound biosynthesized nonenzymatically by a free-radical oxygenation of arachidonic acid, was measured in plasma in order to assess oxidative stress.

Serum low density lipoprotein concentration6 weeks
Plasma insulin concentration6 weeks
Erythrocyte phospholipid fatty acid profiles6 weeks

The EPA and DHA content of erythrocyte lipids were used as a marker of compliance.

Serum adiponectin concentration6 weeks

Trial Locations

Locations (1)

Diabetes & Nutritional Sciences Division, King's College London

🇬🇧

London, United Kingdom

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