Effect of EPA and DHA in the Inflammation and Metabolic Disorders in DMD/DMB Patients

Study Overview
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Brief Summary
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The purpose of this study is to evaluate the effect of docosahexaenoic fatty acid and eicosapentaenoic fatty acid supplementation for six months on the inflammation state as well as the process of muscular regeneration and the metabolic disorders like obesity and insulin resistance in patients with Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (DMB) compared to those receiving placebo.

Detailed Description
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DMD and DMB are X-linked diseases caused by mutations in the DMD gene, these mutations have important functional and structural consequences in skeletal muscle. In muscle fiber is observed inflammation and necrosis as a result of lost regenerative capacity. The muscle fibers can be replaced by connective and adipose tissue. In a previous study the investigators identified that 50% of Duchenne and Becker patients in the range of thirteen years old have obesity. In addition, these patients (N=66) have hyperinsulinemia (53.7%) and insulin resistance (48.5%). It is well known that obesity, hyperinsulinemia and insulin resistance have a inflammatory background. It has been demonstrated that eicosapentaenoic fatty acid (EPA) and docosahexaenoic fatty acid (DHA) exhibit anti-inflammatory properties and have beneficial effects on obesity, hyperinsulinemia and insulin resistance in children and adolescents. Objective: Determine the effect of EPA and DHA on inflammation, obesity and insulin resistance in patients with DMD/DMB compared to those receiving placebo.

Official Title
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Effect of Eicosapentaenoic Fatty Acid (EPA) and Docosahexaenoic Fatty Acids (DHA) Supplementation on the Inflammation State and Metabolic Disorders in Patients With Duchenne Muscular Dystrophy or Becker Muscular Dystrophy

Conditions
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Muscular Dystrophy, Duchenne

Intervention / Treatment
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* Dietary Supplement: EPA and DHA
* Dietary Supplement: Placebo Comparator


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Written informed consent and assent by the patient and both parents or guardian. Patients with clinical diagnosis of Duchenne Muscular Dystrophy (DMD) or Becker Muscular Dystrophy (DMB) Patients were not under treatment with corticosteroids Exclusion Criteria: Patients decided to withdraw from the study Consumption of dietary supplements containing polyunsaturated fatty acids omega 3. With hypersensitivity to fish oil. Patients with respiratory and gastrointestinal problems. Medical responsible assessment the presence of respiratory and gastrointestinal problems. Patients with difficulty swallowing food, including those who have the difficulty ingesting oil capsules. Gastrostomy fed patients.

Ages Eligible for Study
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Minimum Age: 6 Years
Maximum Age: 18 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Prevention
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Quadruple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: EPA and DHA<br>Supplementation of 2.7 g/d of EPA and DHA were provided in 10 capsules per day (4 in the morning, 3 in the afternoon and 3 at night) during a period of 6 months. The capsules sizes were specially for children to improved the feeding process and its presentation is in gelatin capsules. The supplement is purified fish oil with pharmaceutical grade. | Dietary Supplement: EPA and DHA<br>* Each capsule contains 225mg of DHA, 45mg of EPA, other omega 3 fatty acids 20mg.<br>* Other names: omega 3 fatty acid;|
| Placebo Comparator: Placebo Comparator<br>Supplementation of placebo with sunflower fatty at doses of 2.7 g/d were provided in 10 capsules per day (4 in the morning, 3 in the afternoon and 3 at night) during a period of 6 months. The capsules sizes are specially for children to improved the feeding process. This placebo is sunflower oil, so, it did not present anti-inflammatory or insulin sensitivity effects. | Dietary Supplement: Placebo Comparator<br>* Placebo capsules will contain gelatin and sunflower oil. Fatty acid composition is as follows: lauric (C12:0), 0.19%; myristic (C14:0), 0.29%; palmitic (C16:0), 7.59%; palmitoleic (C16:1), 0.25%; stearic (C18:0), 3.49%; oleic (C18:1), 31.08%; linolenic (C18:3), 1.13%; linoleic (C18:2), 55.64%; DHA 0.02%; arachidic (C20:0), 0.30% and arachidonic (C20:4), 0.01%.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Body Composition (Body Fat) | We observed changes in body composition such as total body fat by Dual X-ray Absorptiometry (DXA). | At baseline and at months 3 and 6 of supplementation. | 
| Lean Mass | We observed changes in body composition such as total lean mass by Dual X-ray Absorptiometry (DXA). | At baseline and at months 1, 2, 3, 4, 5 and 6 of supplementation. | 
| Anthropometric Measurement: Body Mass Index | We measured weight, height by anthropometric to calculate the body mass index (body mass index). | At baseline and at months 1, 2, 3, 4, 5 and 6 of supplementation. | 
| Glucose in Serum | A fasting blood sample was taken; serum glucose (mg/dL) levels were measured by the glucose-oxidase method. | At baseline and at months 1, 2, 3, 4, 5 and 6 of supplementation. | 
| Insulin in Blood | A fasting blood sample was taken; insulin was quantified utilizing a commercial kit, that is based on the radioimmunoanalysis method (RIA). | At baseline and at months 1, 2, 3, 4, 5 and 6 of supplementation. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Inflammation Biomarkers (TNF-A) | Plasma cytokine TNF-A was determined by enzyme-linked immunosorbent assay (ELISA) with a multiplex kit in picograms/mL. | Time Frame: At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Inflammation Biomarkers (IL-1) | Plasma cytokine IL-1 was determined by enzyme-linked immunosorbent assay (ELISA) with a multiplex kit in picograms/mL. | Time Frame: At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Inflammation Biomarkers (IL-6) | Plasma cytokine IL-6 was determined by enzyme-linked immunosorbent assay (ELISA) with a multiplex kit in picograms/mL. | Time Frame: At baseline and at months 1, 2, 3, and 6 of supplementation. | 
| Inflammation Biomarkers (IL-10) | Plasma cytokine IL-10 was determined by enzyme-linked immunosorbent assay (ELISA) with a multiplex kit in picograms/mL. | Time Frame: At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Inflammation Biomarker (IL-6 Expression) | The messenger ribonucleic acid (mRNA) expression of cytokines IL-6 from circulating leucocytes was determined by quantifying the real-time polymerase chain reaction (PCR). | Time Frame: At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Inflammation Biomarker (TNF-A Expression) | The messenger ribonucleic acid (mRNA) expression of cytokines TNF-A from circulating leucocytes was determined by quantifying the real-time polymerase chain reaction (PCR) | Time Frame: At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Inflammation Biomarker (IL-1 Expression) | The messenger ribonucleic acid (mRNA) expression of cytokines IL-1 was determined by quantifying the real-time polymerase chain reaction (PCR). | Time Frame: At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Markers of Muscle Degeneration (Creatinine Kinase) | The concentration in serum of CK was determined by chemiluminescent immunometric assay in U/L. | Time Frame: At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Markers of Muscle Degeneration (MMP9) | Plasma matrix metalloproteinase 9 (MMP9) was determined by enzyme-linked immunosorbent assay (ELISA) with a multiplex kit in ng/mL. | Time Frame: At baseline and at months 1, 2, 3 of supplementation. | 
| Markers of Muscle Degeneration (sFas) | The concentration in plasma of soluble Fas (sFas) was determined by enzyme-linked immunosorbent assay (ELISA) with a multiplex kit in picograms/mL. | Time Frame: At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Markers of Muscle Degeneration (Receptor of Fas) | The concentration in plasma of the receptor o Fas (rFas) was determined by enzyme-linked immunosorbent assay (ELISA) with a multiplex kit in picograms/mL. | At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Markers of Muscle Regeneration (VEGF) | Vascular endothelial growth factor (VEGF) was quantified using enzyme linked immunosorbent assay (ELISA). | Time Frame: At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Markers of Muscle Regeneration (FGF) | Plasma marker of regeneration fibroblast growth factor basic (FGF) was determined by enzyme-linked immunosorbent assay (ELISA) with a multiplex kit in picograms/mL. | Time Frame: At baseline and at months 1, 2, 3 and 6 of supplementation. | 
| Incorporation of DHA in the Erythrocytes | The percentage of DHA in the membrane of erythrocytes was determinated by gas chromatography. | Time Frame: At baseline and at months 1, 2, 3, 4, 5 and 6 of supplementation. | 
| Incorporation of EPA in the Erythrocytes | The percentage of EPA in the membrane of erythrocytes was determinated by gas chromatography. | Time Frame: At baseline, at 1, 2, 3, 4, 5, and 6 | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Muscular Dystrophy, Duchenne Muscular Dystrophy, Becker Muscular Dystrophy, Omega 3, Eicosapentaenoic fatty acid, Docosahexaenoic fatty acid