Hypertriglyceridemia, Normal
Conditions
Keywords
cardiovascular risk, omega 3, hydroxytyrosol, hypertriglyceridemia
Brief summary
The primary aim of this study is to investigate the potential cardioprotective effects of daily nutritional doses of eicosapentaenoic acid (EPA) enriched with hydroxytyrosol (HT) on cardiovascular health, as well as its modulatory impact on the intestinal microbiota. This investigation will be conducted through a randomized, double-blind, crossover, and controlled nutritional intervention trial. The study population will include two cohorts: one with normotriglyceridemic individuals, and another with hypertriglyceridemic individuals. Participants will be randomly allocated to one of three intervention groups: (1) EPA+HT supplementation, (2) EPA alone, or (3) a placebo consisting of sunflower oil devoid of EPA.
Detailed description
The product to be tested is a combination of two key components: an oily preparation containing 97% eicosapentaenoic acid (EPA) and a natural extract (Oleacore®) with a high hydroxytyrosol content (10%). Oleacore® is naturally derived from olives using exclusively physical and mechanical methods and is commercialized by Olivenova Health SL (https://olivenova.com/es/). The EPA-97% component is supplied by Beeps Holding GMBH (https://bepsholding.com/el-holding/?lang=es). Three types of nutraceutical formulations will be prepared, and study participants will consume three capsules daily to achieve the following dosages: 1. 2 g of EPA + 15 mg of hydroxytyrosol (HT) (3 Type-1 capsules, EPA+HT), 2. 2 g of EPA (3 Type-2 capsules, EPA), and 3. 2 g of seed oil (sunflower) as a placebo (3 Type-3 capsules, Placebo). The daily dose of 15 mg HT was selected based on the EFSA claim for the prevention of LDL oxidation , while the 2 g EPA dose was established following EFSA recommendations for triglyceride reduction. A chronic, randomized, controlled, crossover, double-blind, free-living study design is proposed, spanning 46 weeks. Following a two-week washout period, during which the consumption of extra virgin olive oil, and olives will be restricted, participants will be grouped based on baseline characteristics (control or hypertriglyceridemia). They will then be randomized into three groups (randomization performed using SPSS software), such that one-third of participants will receive EPA+HT, another third will receive EPA without HT, and the remaining third will receive a placebo nutraceutical containing sunflower oil with no EPA or HT. Each intervention phase will last 12 weeks. After completing one intervention, participants will undergo a four-week washout period before transitioning to the next intervention. Before the start of each phase, participants will be provided with all necessary capsules (EPA+HT, EPA, or placebo) along with detailed consumption guidelines.Volunteers will take the capsules with lunch and dinner. Additionally, during the study, volunteers will consume a commercial olive oil with a minimal phenol content as their culinary oil. This olive oil will also be supplied by the research team for family use (1 L/2 weeks). Participants will be instructed to maintain their usual lifestyle throughout the study but will be restricted from consuming any other culinary oils apart from the one provided by the research group, as well as olives. Furthermore, the use of dietary supplements containing antioxidants, other than vitamins, will be prohibited. Participants will attend the Human Nutrition Unit (UNH) at ICTAN-CSIC on six occasions, corresponding to the beginning and end of each intervention phase. Once eligibility has been confirmed based on the inclusion and exclusion criteria, participants will attend the UNH for signing the informed consent, receiving olive oil for household consumption, and receiving instructions to begin the study. During visits V1-V6 (corresponding to the start and end of each intervention with the different nutraceuticals), participants will report to the UNH at ICTAN-CSIC. Upon arrival, they will provide a fasting urine sample from their first morning void and a stool sample collected within the previous 24 hours. Additionally, a trained nurse will collect a fasting blood sample (40 mL, distributed into tubes containing various anticoagulants). Blood pressure and heart rate will also be measured. Furthermore, participants will undergo a comprehensive anthropometric and body composition assessment. Dietary habits and physical activity levels will be monitored and evaluated during these visits. Compliance with the intervention will be monitored through monthly telephone follow-ups, the collection of unused capsules, and the measurement of specific biomarkers, including plasma phosphatidylcholine-EPA levels as a marker of omega-3 fatty acid intake and urinary HT metabolites.
Interventions
DIETARY_SUPPLEMENTEPA + HT
Volunteers will consume during 12 weeks 2 g/d of EPA and 15 mg/d of HT, corresponding to 3 capsules. Additionally, they will consume a commercial olive oil with a minimal phenol content as their culinary oil that will be suplied by the research team for family use (1L/2 weeks)
DIETARY_SUPPLEMENTEPA
Volunteers will consume during 12 weeks 2 g/d of EPA corresponding to 3 capsules. Additionally, they will consume a commercial olive oil with a minimal phenol content as their culinary oil that will be suplied by the research team for family use (1L/2 weeks)
DIETARY_SUPPLEMENTSunflower Oil
Volunteers will consume during 12 weeks 2 g/d of Sunflower Oil, corresponding to 3 capsules. Additionally, they will consume a commercial olive oil with a minimal phenol content as their culinary oil that will be suplied by the research team for family use (1L/2 weeks)
Sponsors
Beps Holding GmbH
Olivenova Health Sl
National Research Council, Spain
Study design
Allocation
RANDOMIZED
Intervention model
CROSSOVER
Primary purpose
PREVENTION
Masking
DOUBLE (Subject, Investigator)
Intervention model description
Chronic, randomized, controlled, cross-over, double-blind, and free-living study
Eligibility
Sex/Gender
ALL
Age
18 Years to 70 Years
Healthy volunteers
Yes
Inclusion criteria
* Age: 18 to 70 years old * Hypertriglyceridemia (150-250 mg/dl) or normal levels of triglycerides levels (\<150 mg/dl)
Exclusion criteria
* Presence of chronic illnesses * Currently undergoing pharmacological treatment * Smoking * Pregnancy * Adherence to a vegetarian or vegan diet
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Antioxidant enzyme activities (NOX, GSH, GPx, SOD) | 12 months | NADPH oxidase (NOX), glutathione (GSH), glutathione peroxidase (GPx), and superoxide dismutase (SOD) will be measured using ELISA-based methods. |
| Aminotransferases: AST (ASAT) and ALT (ALAT) enzyme activity | 12 months | Measured using enzymatic methods. |
| Uric acid, creatinine and glucose concentrations | 12 months | Measured using standardised spectrophotometric methods. |
| Endothelial biomarkers | 12 months | VCAM-1, ICAM-1, eNOS, E-selectin, and P-selectin concentrations will be measured in serum using ELISA. |
| Platelet function markers | 12 months | Leukotriene B4 and prostaglandin E2 will be measured in serum using ELISAs (Cayman kits). |
| Serum cytokines (IL-6, IFN-γ, TNF-α, IL-10) | 12 months | Serum concentrations of interleukin-6 (IL-6), interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), and interleukin-10 (IL-10) will be determined using ELISA. These biomarkers reflect both pro- and anti-inflammatory responses. |
| C-Reactive Protein (CRP) | 12 months | CRP will be measured using an automated ultrasensitive turbidimetric method (AU2700 Biochemistry analyser, Olympus). |
| Oxidized LDL and 8-hydroxy-2'-deoxyguanosine (8-OHdG) | 12 months | LDLox and 8-OHdG will be measured in serum using ELISA. |
| Lipid peroxidation (TBARS) | 12 months | TBARS will be measured as µM MDA equivalents using ELISA. |
| TAS will be assessed using a colorimetric assay or ELISA. | 12 months | TAS will be assessed using a colorimetric assay or ELISA. |
| Total oxidative status (TOS) | 12 months | TOS will be measured using ELISA. |
| Total cholesterol concentration | 12 months | Total cholesterol in serum samples will be determined following reference methods recommended by the Sociedad Española de Bioquímica Clínica y Patología Molecular (SEQC) using the Roche Cobas Integra 400 plus analyser (Roche Diagnostics). |
| HDL-C, LDL-C, VLDL-C, and triglyceride concentrations | 12 months | Measured in serum using reference or enzymatic methods recommended by SEQC. |
| Apolipoprotein A1 and B concentrations | 12 months | Measured with clinical chemistry methods based on SEQC guidelines. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Resting Heart Rate | 12 months | Heart rate will be measured using the same automatic arm sphygmomanometer (BS 150; Pic Indolor Diagnostic) under the same conditions as blood pressure. Volunteers will rest for 5 minutes prior to the measurement. If multiple readings are required due to variation, the mean of the two (or three) closest values will be used. Heart rate will be expressed in beats per minute (bpm). |
| Fecal Short Chain Fatty Acids (SCFAs) Concentration | 12 months | Short chain fatty acids (SCFAs) determination in faeces.The analytes will be analysed by gas chromatography system (Agilent Technologies Palo Alto, CA, USA) equipped with a mass spectrometer (MS) detector and Agilent DB-WAXtr column (100% polyethilen glycol, 60 m, 0.325 mm, 0250 um). |
| Fecal Lactate Concentration | 12 months | Lactate levels in fecal samples will be measured using an Advanced Compact Ion Chromatographic instrument IC867 (Metrohm AG, Herisau, Switzerland). |
| Gut Microbiota Composition (16S rRNA Gene Profiling) | 12 months | Microbial composition of fecal samples will be assessed by 16S rRNA gene amplicon sequencing. Genomic DNA will be extracted using the Quick-DNA™ Fecal/Soil Microbe Miniprep Kit (Zymo Research). Library preparation and sequencing will be conducted at Maastricht University (the Netherlands). Data analysis will be performed using the QIIME software package (v1.9.0), with taxonomic assignment based on the Greengenes database (v13.8). |
| Fasting glucose | 12 months | Measured using SEQC-recommended methods. |
| Fasting insulin and C-peptide | 12 months | Measured using immunoassays or ELISA. |
| Glycated hemoglobin (HbA1c) | 12 months | Measured using HPLC or immunoturbidimetric methods. |
| Incretins (Glucagon and GLP-1) | 12 months | Glucagon and GLP-1 concentrations will be measured using ELISA. |
| Adipokines (Leptin and PAI-1) | 12 months | Leptin and plasminogen activator inhibitor-1 (PAI-1) will be measured using ELISA. |
| Systolic and Diastolic Blood Pressure | 12 months | Systolic and diastolic blood pressure will be measured using an automatic arm sphygmomanometer (BS 150; Pic Indolor Diagnostic) at baseline and at the end of each intervention period. Measurements will be taken after the volunteer has rested on a chair for 5 minutes. Two consecutive readings will be taken on the left arm. If the values differ by more than 10-15 mmHg, a third reading will be recorded. The mean of the two (or three) closest readings will be used for analysis. Blood pressure will be expressed in mmHg.. |
Countries
Spain
Outcome results
None listed