Healthy, Hyperlipidemia
Conditions
Keywords
Hyperlipidemia, coronary heart disease, gene expression, fish oil, DHA, EPA
Brief summary
The aim of this study is to investigate the effects of short- and long-term intervention with EPA and DHA-rich fish oil on gene expression profiles in healthy and hyperlipidemic males.
Detailed description
Cardiovascular and coronary heart diseases continue to be the leading causes of morbidity and mortality among adults in Europe and North America. Since the number of elderly people and therefore the number of chronic-inflammatory diseases rise, preventive therapies become more important. Within preventive strategies, nutrition plays a central role. Cross-sectional studies suggested that omega-3 fatty acids, especially the very long-chain fatty acids Eicosapentaenoic acid (EPA, C20:5ω3) and Docosahexaenoic acid (DHA, C22:6ω3), are protective against cardiovascular and coronary heart diseases. Their cardio protective potential is based on their positive effects on blood lipids, vascular tonus and blood clotting. A number of controlled clinical trials have shown that EPA and DHA supplementation lower fasting and postprandial plasma concentrations of triglyceride-rich lipoproteins and their remnants. Biochemical research revealed numerous metabolic effects of EPA and DHA, ranging from their effects on membrane fluidity to the modification of the eicosanoid profile. However, only a few human clinical trials examined the regulative effects of DHA and EPA supplementation on gene expression. Furthermore, to our knowledge no published research data is available dealing with the effect of these fatty acids on gene expression in subjects with hypertriglyceridemia in comparison to healthy subjects. Such findings are of great concern due to hints that especially people with hypertriglyceridemia benefit from the triglyceride lowering effect of EPA and DHA supplementation. Presently it is not well-established if the gene regulative potential of EPA and DHA in these persons differs from healthy persons. These findings could help to understand the differences in the metabolic effects of EPA and DHA in healthy vs. hypertriglyceridemic persons, which have a greater risk for cardiovascular and coronary diseases. Finally, these data could contribute to a knowledge basis for targeted strategies in preventive therapies with the very long-chain omega-3 fatty acids EPA and DHA.
Interventions
DIETARY_SUPPLEMENTFish oil
Dietary Supplement: fish oil capsules (6 per day) 3024 mg n-3 fatty acids daily (1512 mg EPA and 1008 mg DHA) about 3 months
DIETARY_SUPPLEMENTPlacebo (corn oil)
corn oil (6 capsules per day)
Sponsors
Gottfried Wilhelm Leibniz Universität Hannover
Study design
Allocation
RANDOMIZED
Intervention model
PARALLEL
Primary purpose
PREVENTION
Masking
DOUBLE (Subject, Investigator)
Eligibility
Sex/Gender
MALE
Age
20 Years to 51 Years
Healthy volunteers
Yes
Inclusion criteria
* males, 20-50 years * non-smokers * ethnicity: Caucasians * no medical treatment * healthy subjects: * no documented disease * normal blood lipids (triglyceride \< 150 mg/dl; total cholesterol \< 200 mg/dl) * humans with increased blood lipids (hyperlipidemia) * documented hypertriglyceridemia or * triglyceride ≥ 150 mg/dl (≥ 1,7 mmol/l) and * total cholesterol \> 200 mg/dl (5,2 mmol/l) * written confirmation of the subjects after detailed oral and written explanation about the study contents, - requirements and risks * ability and willingness of the participants to attend the investigator's orders (compliance of the study conditions, consumption of the study medicaments according to the dosage commendation)
Exclusion criteria
* Body-Mass-Index (BMI) ≥ 35 * smokers * medical treatment (especially corticosteroids, anti-inflammatory drugs, blood lipids lowering drugs (e.g. statins, fibrates, bile acid exchanger resin, phytosterols) * taking any supplements with omega-3 fatty acids, phytosterols, polyglucosamines (Chitosan) or other lipid binding ingredients * daily consumption of omega-3 fatty acids rich fish (salmon, mackerel, herring) * heavy chronic diseases (tumors, diabetes typ 1, etc.), documented heart disease, documented blood clotting disorders, renal failure, liver diseases * documented blood clotting disorders and consumption of coagulation-inhibiting drugs (for example Marcumar, ASS) * allergy or intolerance to fish/fish oil or any of the study ingredients of the test products * chronic gastro-intestinal diseases (Colitis ulcerosa, Morbus Crohn, pancreatic insufficiency) * donation of blood in the last 6 weeks * routine consumption of laxative * common
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Gene Expression Changes | Gene expression changes (number of regulated genes) | Gene expression changes were measured by using whole genome microarrays. The expression values of all genes were compared between baseline and 4 hours, 7 days and twelve weeks after supplementation with FO or CO and differentially expressed genes were detected by standard two-state pooled-variance t-test (p\<0,05). The number of differentially expressed genes (regulated genes)compared to the baseline values were determined for every study group in total as well as for every time point (4 hours, 7 days, 12 weeks)in total and specifically. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Fatty Acid Composition of Erythrocyte Membranes (Omega-3 Index) | baseline and after 12 weeks | Fasting venous blood samples were collected and RBC membrane FA composition including the omega-3 index, given as EPA + DHA, was analyzed at baseline and after 12 weeks according to the omega-3 index methodology (Harris & von Schacky, 2004). Results are presented as a percentage of the total identified FAs after response factor correction. The coefficient of variation for EPA + DHA was 5%. Quality was assured according to DIN ISO 15189. |
| Blood Lipids | baseline and after 12 weeks | Fasting venous blood samples were collected and blood lipid levels were determined by an external contract laboratory (LADR, Hannover; Germany) at baseline (t0), after one week (t1) and after 12 weeks (t12) of supplementation. |
Countries
Germany
Participant flow
Recruitment details
The recruitment of subjects was performed by several advertisements and study placards in hanover from january until february 2010. By a telephonic preselection 106 subjects were selected according to inclusion criteria.
Pre-assignment details
Serum lipid levels of the preselected subjects were determined and 20 normolipidemic and 20 dyslipidemic men were enrolled in the study population.
Participants by arm
| Arm | Count |
|---|---|
| Placebo - Healthy Subjects Dietary Supplement: corn oil capsules (6 per day) about 3 months n=6 | 10 |
| Placebo - Hyperlipedemic Subjects Dietary Supplement: corn oil capsules (6 per day) about 3 months n=8 | 10 |
| Fish Oil - Hyperlipidemic Subjects Dietary Supplement: fish oil capsules (6 per day) 3024 mg n-3 fatty acids daily (1512 mg EPA and 1008 mg DHA) about 3 months n=8 | 10 |
| Fish Oil - Healthy Subjects Dietary Supplement: fish oil capsules (6 per day) 3024 mg n-3 fatty acids daily (1512 mg EPA and 1008 mg DHA) about 3 months. n=9 | 10 |
| Total | 40 |
Baseline characteristics
| Characteristic | Placebo - Hyperlipedemic Subjects | Fish Oil - Hyperlipidemic Subjects | Placebo - Healthy Subjects | Fish Oil - Healthy Subjects | Total |
|---|---|---|---|---|---|
| Age, Categorical <=18 years | 0 Participants | 0 Participants | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical >=65 years | 0 Participants | 0 Participants | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical Between 18 and 65 years | 10 Participants | 10 Participants | 10 Participants | 10 Participants | 40 Participants |
| Age Continuous | 41.8 years STANDARD_DEVIATION 8.94 | 40.2 years STANDARD_DEVIATION 8.64 | 37.4 years STANDARD_DEVIATION 8.3 | 37.5 years STANDARD_DEVIATION 8.11 | 38.82 years STANDARD_DEVIATION 8.29 |
| Region of Enrollment Germany | 10 participants | 10 participants | 10 participants | 10 participants | 40 participants |
| Sex: Female, Male Female | 0 Participants | 0 Participants | 0 Participants | 0 Participants | 0 Participants |
| Sex: Female, Male Male | 10 Participants | 10 Participants | 10 Participants | 10 Participants | 40 Participants |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk | EG002 affected / at risk | EG003 affected / at risk |
|---|---|---|---|---|
| deaths Total, all-cause mortality | — / — | — / — | — / — | — / — |
| other Total, other adverse events | 0 / 6 | 0 / 8 | 0 / 8 | 0 / 9 |
| serious Total, serious adverse events | 0 / 6 | 0 / 8 | 0 / 8 | 0 / 9 |
Outcome results
Primary
Gene Expression Changes
Gene expression changes were measured by using whole genome microarrays. The expression values of all genes were compared between baseline and 4 hours, 7 days and twelve weeks after supplementation with FO or CO and differentially expressed genes were detected by standard two-state pooled-variance t-test (p\<0,05). The number of differentially expressed genes (regulated genes)compared to the baseline values were determined for every study group in total as well as for every time point (4 hours, 7 days, 12 weeks)in total and specifically.
Time frame: Gene expression changes (number of regulated genes)
| Arm | Measure | Group | Value (NUMBER) |
|---|---|---|---|
| Placebo - Healthy Subjects | Gene Expression Changes | total (4 hours+7 days+12weeks,without doubles) | 841 number of regulated genes |
| Placebo - Healthy Subjects | Gene Expression Changes | 7 days (specific) | 135 number of regulated genes |
| Placebo - Healthy Subjects | Gene Expression Changes | 4 hours (specific) | 94 number of regulated genes |
| Placebo - Healthy Subjects | Gene Expression Changes | 4 hours (total) | 320 number of regulated genes |
| Placebo - Healthy Subjects | Gene Expression Changes | 12 weeks (specific) | 312 number of regulated genes |
| Placebo - Healthy Subjects | Gene Expression Changes | 7 days (total) | 418 number of regulated genes |
| Placebo - Healthy Subjects | Gene Expression Changes | 12 weeks (total) | 500 number of regulated genes |
| Placebo - Hyperlipedemic Subjects | Gene Expression Changes | 7 days (specific) | 242 number of regulated genes |
| Placebo - Hyperlipedemic Subjects | Gene Expression Changes | 12 weeks (total) | 493 number of regulated genes |
| Placebo - Hyperlipedemic Subjects | Gene Expression Changes | 7 days (total) | 629 number of regulated genes |
| Placebo - Hyperlipedemic Subjects | Gene Expression Changes | 4 hours (specific) | 85 number of regulated genes |
| Placebo - Hyperlipedemic Subjects | Gene Expression Changes | 12 weeks (specific) | 287 number of regulated genes |
| Placebo - Hyperlipedemic Subjects | Gene Expression Changes | 4 hours (total) | 436 number of regulated genes |
| Placebo - Hyperlipedemic Subjects | Gene Expression Changes | total (4 hours+7 days+12weeks,without doubles) | 1041 number of regulated genes |
| Fish Oil - Hyperlipidemic Subjects | Gene Expression Changes | 12 weeks (total) | 1027 number of regulated genes |
| Fish Oil - Hyperlipidemic Subjects | Gene Expression Changes | total (4 hours+7 days+12weeks,without doubles) | 1639 number of regulated genes |
| Fish Oil - Hyperlipidemic Subjects | Gene Expression Changes | 4 hours (total) | 508 number of regulated genes |
| Fish Oil - Hyperlipidemic Subjects | Gene Expression Changes | 7 days (total) | 1105 number of regulated genes |
| Fish Oil - Hyperlipidemic Subjects | Gene Expression Changes | 4 hours (specific) | 246 number of regulated genes |
| Fish Oil - Hyperlipidemic Subjects | Gene Expression Changes | 7 days (specific) | 326 number of regulated genes |
| Fish Oil - Hyperlipidemic Subjects | Gene Expression Changes | 12 weeks (specific) | 203 number of regulated genes |
| Fish Oil - Healthy Subjects | Gene Expression Changes | 7 days (total) | 230 number of regulated genes |
| Fish Oil - Healthy Subjects | Gene Expression Changes | 12 weeks (specific) | 214 number of regulated genes |
| Fish Oil - Healthy Subjects | Gene Expression Changes | 7 days (specific) | 78 number of regulated genes |
| Fish Oil - Healthy Subjects | Gene Expression Changes | 4 hours (total) | 264 number of regulated genes |
| Fish Oil - Healthy Subjects | Gene Expression Changes | total (4 hours+7 days+12weeks,without doubles) | 628 number of regulated genes |
| Fish Oil - Healthy Subjects | Gene Expression Changes | 4 hours (specific) | 140 number of regulated genes |
| Fish Oil - Healthy Subjects | Gene Expression Changes | 12 weeks (total) | 338 number of regulated genes |
Secondary
Blood Lipids
Fasting venous blood samples were collected and blood lipid levels were determined by an external contract laboratory (LADR, Hannover; Germany) at baseline (t0), after one week (t1) and after 12 weeks (t12) of supplementation.
Time frame: baseline and after 12 weeks
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Placebo - Healthy Subjects | Blood Lipids | total cholesterol (TC) baseline | 194.0 mg/dl | Standard Deviation 22.2 |
| Placebo - Healthy Subjects | Blood Lipids | total cholesterol (TC) after 12 weeks | 188.9 mg/dl | Standard Deviation 21.5 |
| Placebo - Healthy Subjects | Blood Lipids | low-density lipoprotein (LDL) baseline | 118.1 mg/dl | Standard Deviation 25.1 |
| Placebo - Healthy Subjects | Blood Lipids | low-density lipoprotein (LDL) after 12 weeks | 118.6 mg/dl | Standard Deviation 28.2 |
| Placebo - Healthy Subjects | Blood Lipids | high-density lipoprotein (HDL) baseline | 52.7 mg/dl | Standard Deviation 12.7 |
| Placebo - Healthy Subjects | Blood Lipids | high-density lipoprotein (HDL) after 12 weeks | 52.6 mg/dl | Standard Deviation 11.6 |
| Placebo - Healthy Subjects | Blood Lipids | triacylglycerol (TG) baseline | 115.6 mg/dl | Standard Deviation 53.8 |
| Placebo - Healthy Subjects | Blood Lipids | triacylglycerol (TG) after 12 weeks | 87.4 mg/dl | Standard Deviation 14.8 |
| Placebo - Hyperlipedemic Subjects | Blood Lipids | total cholesterol (TC) after 12 weeks | 257.5 mg/dl | Standard Deviation 60.3 |
| Placebo - Hyperlipedemic Subjects | Blood Lipids | high-density lipoprotein (HDL) baseline | 47.9 mg/dl | Standard Deviation 10 |
| Placebo - Hyperlipedemic Subjects | Blood Lipids | triacylglycerol (TG) baseline | 175.0 mg/dl | Standard Deviation 56.2 |
| Placebo - Hyperlipedemic Subjects | Blood Lipids | high-density lipoprotein (HDL) after 12 weeks | 47.4 mg/dl | Standard Deviation 8.21 |
| Placebo - Hyperlipedemic Subjects | Blood Lipids | triacylglycerol (TG) after 12 weeks | 186.9 mg/dl | Standard Deviation 49.6 |
| Placebo - Hyperlipedemic Subjects | Blood Lipids | low-density lipoprotein (LDL) after 12 weeks | 172.6 mg/dl | Standard Deviation 54.4 |
| Placebo - Hyperlipedemic Subjects | Blood Lipids | low-density lipoprotein (LDL) baseline | 178.1 mg/dl | Standard Deviation 45.4 |
| Placebo - Hyperlipedemic Subjects | Blood Lipids | total cholesterol (TC) baseline | 261.0 mg/dl | Standard Deviation 48.1 |
| Fish Oil - Hyperlipidemic Subjects | Blood Lipids | triacylglycerol (TG) baseline | 322.4 mg/dl | Standard Deviation 259 |
| Fish Oil - Hyperlipidemic Subjects | Blood Lipids | low-density lipoprotein (LDL) baseline | 146.1 mg/dl | Standard Deviation 5.31 |
| Fish Oil - Hyperlipidemic Subjects | Blood Lipids | low-density lipoprotein (LDL) after 12 weeks | 167.4 mg/dl | Standard Deviation 23.6 |
| Fish Oil - Hyperlipidemic Subjects | Blood Lipids | triacylglycerol (TG) after 12 weeks | 230.8 mg/dl | Standard Deviation 147 |
| Fish Oil - Hyperlipidemic Subjects | Blood Lipids | high-density lipoprotein (HDL) baseline | 45.2 mg/dl | Standard Deviation 6.5 |
| Fish Oil - Hyperlipidemic Subjects | Blood Lipids | high-density lipoprotein (HDL) after 12 weeks | 50.8 mg/dl | Standard Deviation 9.43 |
| Fish Oil - Hyperlipidemic Subjects | Blood Lipids | total cholesterol (TC) baseline | 262.2 mg/dl | Standard Deviation 61.9 |
| Fish Oil - Hyperlipidemic Subjects | Blood Lipids | total cholesterol (TC) after 12 weeks | 264.4 mg/dl | Standard Deviation 49.2 |
| Fish Oil - Healthy Subjects | Blood Lipids | triacylglycerol (TG) after 12 weeks | 66.2 mg/dl | Standard Deviation 16.7 |
| Fish Oil - Healthy Subjects | Blood Lipids | low-density lipoprotein (LDL) after 12 weeks | 117.2 mg/dl | Standard Deviation 22.1 |
| Fish Oil - Healthy Subjects | Blood Lipids | total cholesterol (TC) after 12 weeks | 198.6 mg/dl | Standard Deviation 26.1 |
| Fish Oil - Healthy Subjects | Blood Lipids | total cholesterol (TC) baseline | 184.1 mg/dl | Standard Deviation 13.3 |
| Fish Oil - Healthy Subjects | Blood Lipids | triacylglycerol (TG) baseline | 82.4 mg/dl | Standard Deviation 35.3 |
| Fish Oil - Healthy Subjects | Blood Lipids | high-density lipoprotein (HDL) baseline | 59.0 mg/dl | Standard Deviation 10.4 |
| Fish Oil - Healthy Subjects | Blood Lipids | low-density lipoprotein (LDL) baseline | 108.7 mg/dl | Standard Deviation 12.8 |
| Fish Oil - Healthy Subjects | Blood Lipids | high-density lipoprotein (HDL) after 12 weeks | 65.2 mg/dl | Standard Deviation 14.4 |
Secondary
Fatty Acid Composition of Erythrocyte Membranes (Omega-3 Index)
Fasting venous blood samples were collected and RBC membrane FA composition including the omega-3 index, given as EPA + DHA, was analyzed at baseline and after 12 weeks according to the omega-3 index methodology (Harris & von Schacky, 2004). Results are presented as a percentage of the total identified FAs after response factor correction. The coefficient of variation for EPA + DHA was 5%. Quality was assured according to DIN ISO 15189.
Time frame: baseline and after 12 weeks
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Placebo - Healthy Subjects | Fatty Acid Composition of Erythrocyte Membranes (Omega-3 Index) | omega-3 index baseline | 5.08 percentage of total fatty acids | Standard Deviation 1.41 |
| Placebo - Healthy Subjects | Fatty Acid Composition of Erythrocyte Membranes (Omega-3 Index) | omega-3 index after 12 weeks | 4.78 percentage of total fatty acids | Standard Deviation 1.16 |
| Placebo - Hyperlipedemic Subjects | Fatty Acid Composition of Erythrocyte Membranes (Omega-3 Index) | omega-3 index after 12 weeks | 5.50 percentage of total fatty acids | Standard Deviation 1.57 |
| Placebo - Hyperlipedemic Subjects | Fatty Acid Composition of Erythrocyte Membranes (Omega-3 Index) | omega-3 index baseline | 5.27 percentage of total fatty acids | Standard Deviation 1.69 |
| Fish Oil - Hyperlipidemic Subjects | Fatty Acid Composition of Erythrocyte Membranes (Omega-3 Index) | omega-3 index baseline | 4.90 percentage of total fatty acids | Standard Deviation 1.8 |
| Fish Oil - Hyperlipidemic Subjects | Fatty Acid Composition of Erythrocyte Membranes (Omega-3 Index) | omega-3 index after 12 weeks | 9.84 percentage of total fatty acids | Standard Deviation 1.26 |
| Fish Oil - Healthy Subjects | Fatty Acid Composition of Erythrocyte Membranes (Omega-3 Index) | omega-3 index baseline | 5.24 percentage of total fatty acids | Standard Deviation 0.7 |
| Fish Oil - Healthy Subjects | Fatty Acid Composition of Erythrocyte Membranes (Omega-3 Index) | omega-3 index after 12 weeks | 10.7 percentage of total fatty acids | Standard Deviation 1.06 |