Effects of n-3 Intake on Lipid Profile, Biochemical and Inflammatory Markers in Subjects With Obesity

Study Overview
=================
Brief Summary
-----------------
Obesity is a chronic disease characterized by the excessive accumulation of adipose tissue that is harmful to health. Dietary habits modification through a caloric restriction, macronutrient distribution including linoleic and linolenic (n-6:n-3) polyunsaturated fatty acids (PUFAs) ratio, has been suggested for obesity management. It has been proposed that the optimal n-6: n-3 ratio should be between 1:1 and 5:1 to maintain a healthy balance. Purpose: Compare the effect of a diet n6:n3 ratio of polyunsaturated fatty acids with a group supplemented with n-3 PUFA on lipid profile in erythrocyte membrane, biochemical and inflammatory markers in subjects with obesity. Methods: 58 subjects were randomly divided into two groups: fish oil group and the placebo group. Anthropometric and biochemical data were evaluated, cytokine levels was performed using the Bio-PlexPro™ HumanTh17Cytokine Assays (MagPix) panel. The fatty acid profile quantification in the erythrocyte membrane was carried out by gas chromatography. Statistical analysis was performed with Statistical Package for the Social Sciences (SPSS) v.22 software.

Detailed Description
-----------------
A randomized clinical trial was conducted at the Institute of Translational Nutrigenetics and Nutrigenomics of the University of Guadalajara. All participants were recruited through flyers and social media invitations. Sample size was determined according to the mean difference formula for clinical trials. To achieve a statistical power of 80% and an alpha of 5%, a sample size of 19 participants in each study group was required. However, 58 obese individuals who met the selection criteria were randomized in n-3 group or placebo group. Blood sample, height and weight were measured after 8-12 hour fast and wearing light clothes. The nutritional intervention consisted in 20% reduction by the total estimated energy through the Mifflin-St.Jeor formula was calculated, following a 50% to carbohydrates, 20% to proteins and 30% to lipids distribution. All participants received a balanced nutritional plan of omega-6/omega-3 close to 5:1 ratio, according to estimated energy. Additionally, a high omega-3 foods list was proportioned to emphasize their consumption during the study. Fish oil group besides the diet was supplemented with omega 3, the dosage was 2 capsules per day, containing 1.5 g of total omega 3, of which 1000 mg were EPA and 500mg DHA. The omega 3 capsules were obtained from the same batch, and a toxicity analysis was performed to verify the safety or the placebo group (2 capsules per day made from sunflower oil). This study was approved by the Ethics and Biosafety Committee of the Health Sciences Center, University of Guadalajara (Registration number CI-01219) and was carried out according to the Declaration of Helsinki (2013) and all the participants signed a written consent-informed.

Official Title
-----------------
Effect of a Diet Supplemented With Omega-3 on Biochemical Variables, Cytokine Concentration and Fatty Acid Profile in Subjects With Obesity

Conditions
-----------------
Obesity

Intervention / Treatment
-----------------
* Dietary Supplement: n-3 supplementation group
* Dietary Supplement: Placebo group


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Subjects of both sexes Mestizo from West Mexico Age between 30 to 50 years of age Sign the Informed Consent Diagnostic of Obesity type I and II according to BMI (30 - 40kg / m2) Waist circumference (WC) women ≥80cm, men ≥90cm Sedentary lifestyle ˂ 150 minutes per week Exclusion Criteria: Pregnant or breastfeeding women Diabetes disease Cardiovascular disease Any type of cancer disease Tobacco and alcohol (consumption ≥ 40 g of alcohol per day for men and ≥ 20 g for women) Participants that consume n-3 supplements, anti-inflammatory medications, or some type of lipid-lowering drugs in the past year

Ages Eligible for Study
-----------------
Minimum Age: 25 Years
Maximum Age: 50 Years

Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
Accepts Healthy Volunteers

Study Plan
=================
How is the study designed?
-----------------

Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: Randomized Parallel with placebo
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: n-3 supplementation group<br>The nutritional strategy consisted a hypocaloric diet high in n-3. A 20% reduction by the total estimated energy through the Mifflin-St.Jeor formula was calculated, following a 50% to carbohydrates, 20% to proteins and 30% to lipids distribution. All participants received a balanced nutritional plan of omega-6/omega-3 close to 5:1 ratio, according to estimated energy. Additionally, a high omega-3 foods list was proportioned to emphasize their consumption during the study plus Omega 3 supplementation, the dosage was 2 capsules per day, containing 1.5 g of omega 3, of which 1000 mg were EPA and 500mg DHA. The omega 3 capsules were obtained from the same batch. | Dietary Supplement: n-3 supplementation group<br>* n-3 supplementation group (1.5g of omega 3)<br>|
| Placebo Comparator: Placebo group<br>The nutritional strategy consisted a hypocaloric diet high in n-3. A 20% reduction by the total estimated energy through the Mifflin-St.Jeor formula was calculated, following a 50% to carbohydrates, 20% to proteins and 30% to lipids distribution. All participants received a balanced nutritional plan of omega-6/omega-3 close to 5:1 ratio, according to estimated energy. Additionally, a high omega-3 foods list was proportioned to emphasize their consumption during the study plus placebo capsules (2 capsules per day made from sunflower oil) | Dietary Supplement: Placebo group<br>* Placebo group (sunflower oil)<br>* Other names: Placebo group (sunflower oil);|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| The fatty acid profile quantification | All the samples was performed in a Gas Chromatograph (GC) Agilent Technologies, 6850 network system coupled to an injector (Agilent Technologies, 7083 Series) with a column for fatty acids (Durabond, DB-23). flame ionization detector with helium as gas carrier (0.7 cm3 min-1) and a temperature ramp (110 °C - 220 °C). | Mean change from baseline (0 Month) to end of treatment at 4th Month | 
| Pro-inflammatory and Anti-inflammatory Cytokines profile in serum | The pro-inflammatory and anti-inflammatory cytokines quantification were using Bio-Plex Pro™ Human cytokine Standard 17-Plex, Group I kit following the supplier's instructions, and the read was immediately by MAGPIX™ analyzer. The 17 cytokine include: IL1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12p70, IL-13, IL-15, IL17A, G-CSF, GM-CSF, INF-γ, MCP-1, MIP1α, TNFα. Units of measure in pg/ml. | Mean change from baseline (0 Month) to end of treatment at 4th Month | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in Weight | The weight was measured in kilograms on InBody 370 | At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month | 
| Changes in Body Mass Index (BMI) | Weight and height were be combined to report BMI in kg/m^2 | At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month | 
| Changes in Waist Circumference | Waist circumference was measured at the narrowest point between the edge of the inner rib and the iliac crest, with the participant in an abducted and relaxed position, after expiration using a Lufkin Executive® tape. | At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month. | 
| Changes in Fat Mass | The Fat Mass was measured in kilograms by electrical bioimpedance on InBody 370. | At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month | 
| Changes in Serum Glucose | It was measured in mg/dL using a dry chemistry system in Vitros 350 equipment. | At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month | 
| Changes in Serum Insulin | Were determined through Insulin Model ELISA kit following the supplier's instructions. | At the baseline (0 Month) and the 4th month | 
| Changes in homeostatic model assessment - insulin resistance (HOMA-IR) | Serum glucose and Insulin levels were be combined to report HOMA-IR calculated as described by Matthews. | At the baseline (0 Month) and the 4th month | 
| Changes in Total Cholesterol | It was measured in mg/dL using a dry chemistry system in Vitros 350 equipment. | At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month | 
| Changes in High-density lipoprotein (c-HDL) | It was measured in mg/dL using a dry chemistry system in Vitros 350 equipment. | At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month | 
| Changes in Low-density lipoprotein (c-LDL) | It was calculated using Friedewald formula | At the baseline (0 Month) and 1st month, 2nd month, 3th month and the 4th month | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
Dietary Supplementations, Anti-Inflammatory Agent