Effects of Dietary Amino Acids on Serum and Macrophage Atherogenicity

Study Overview
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Brief Summary
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Recently, the investigators have been screening for anti-atherogenic or pro-atherogenic amino acids (AAs) in the macrophage model system to better understand their role in atherogenesis. The findings so far suggest that specific AAs induce selective anti-atherogenic effects (glycine, alanine, leucine and cysteine) or pro-atherogenic effects (glutamate and glutamine) in macrophages. Taking together the above previous reports with the mechanisms behind macrophage foam cell formation and atherogenesis, it is possible that AAs could be anti-atherogenic or pro-atherogenic via their mechanism of action on macrophage foam cell formation. This paradigm may serve as a basis for the development of novel cardio-protective, anti-atherogenic nutritional, or therapeutic approaches, that should be studied in human trials.

Detailed Description
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Atherosclerosis is the underlying cause of cardiovascular diseases (CVD), the major cause of death worldwide. Atherosclerosis is an inflammatory disease of the arteries in which activated macrophages are abundant in the atherosclerotic lesions. Macrophages play key roles during early atherogenesis. After differentiating from peripheral blood monocytes, the formed intimal macrophages take up oxidized/modified lipoproteins and are transformed into lipid-rich foam cells, the hallmark feature of early atherogenesis. In addition to lipoprotein uptake, lipid accumulation in macrophages can also result from alterations in cellular lipid metabolism, e.g. attenuated reverse lipid transport or enhanced rates of lipid biosynthesis. Although much progress has been made in understanding the role of different lipids (fatty acids, cholesterol, phospholipids or triglycerides) in macrophage foam-cell formation and atherosclerosis development, little is known about the potential impact of other nutrients, such as amino acids (AAs). Previous studies have demonstrated an association between specific AAs and increased CVD risk or higher prevalence of coronary artery disease (CAD). For instance, the role of the branched chain AAs (BCAAs; valine, leucine, and isoleucine) in atherogenesis and CVD has been recently studied. Association studies showed that the plasma levels of BCAAs significantly and independently correlated with dyslipidemia and CAD. However, BCAAs were found to possess cardio-protective effects in a heart failure rat model. Specifically, leucine was reported to attenuate atherosclerosis development in the atherosclerotic apoE-deficient (apoE-/-) mice model, by improving the plasma lipid profile and by reducing systemic inflammation. Other studies have indicated the ability of some AAs to exert anti-atherogenic effects. Glycine, the simplest AA, was inversely associated with the risk of acute myocardial infarction (AMI) in patients with high apolipoprotein B (apoB) and LDL-cholesterol levels. Cysteine is an essential AA in the biosynthesis of glutathione, a key endogenous anti-oxidant known for its anti-atherogenic properties in macrophage lipid metabolism leading to the attenuation of atherosclerosis development. Cysteine analogues such as N-acetyl cysteine (NAC) or ribose cysteine were shown to be anti-atherogenic. Arginine, a basic AA, is the main precursor for nitric oxide production in the vascular endothelium. Arginine was shown to protect endothelial cells from lipid peroxidation, and to delay or reduce atheroma formation. Arginine anti-atherogenicity includes its anti-oxidant properties and its ability to improve endothelial function in CVD or overweight patients. The aim of the proposed study is to investigate the effects of one month of supplementation with specific AAs on changes in the macrophage atherogenicity and lipid metabolism together with other risk markers of atherosclerosis development, such as serum oxidative status and lipid levels, on healthy subjects. Findings from the current proposed study may shed light on yet unknown mechanisms by which specific AAs affect atherosclerosis development and CVD risk and hence could possibly assist in the future development of anti-atherogenic strategies

Official Title
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Effects of Dietary Amino Acids on Serum and Macrophage Atherogenicity

Conditions
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Atherosclerosis, Diet Modification, Serum; Disease

Intervention / Treatment
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* Other: Cellulose (control)
* Dietary Supplement: Glycine
* Dietary Supplement: Alanine
* Dietary Supplement: Leucine
* Dietary Supplement: Isoleucine
* Dietary Supplement: Valine
* Dietary Supplement: Cysteine
* Dietary Supplement: Arginine
* Dietary Supplement: Methionine
* Dietary Supplement: Glutamate
* Dietary Supplement: Glutamine


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: healthy adult males between the ages of 18-50 after signing informed consent Exclusion Criteria: pregnancy, renal, pulmonary, cardiovascular or endocrinal diseases, diabetes, cancer, morbid obesity (body mass index, BMI > 40 kg/m2), heavy smoking (> 20 cigarettes/day), or consumption of more than two alcoholic drinks per day. Subjects from the same hospital unit or from the laboratory where this study is conducted are also excluded

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 50 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Other
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Sham Comparator: Control<br>Other: Cellulose (control): cellulose, water soluble powder, 2 g in one daily dose, during 30 days | Other: Cellulose (control)<br>* Cellulose (a vegetal product, in powder form, used as dietary fiber, that is not digested or absorbed in the intestines), will be supplemented in the form of water soluble powder.<br>|
| Experimental: Glycine (Trade name: Glycine)<br>Dietary Supplement: Glycine, water soluble powder, 7.8 g daily in one dose, during 30 days. | Dietary Supplement: Glycine<br>* Amino acids are the building blocks of proteins, they are essential organic compounds in the normal diet, and are also individually administered as nutritional supplements. Glycine will be supplied in the form of water soluble powder (the commercial form), for one daily dose. The corresponding amino acid doses were established based on the highest intakes at the 99th percentile in adult men (Institute of Medicine. 2005. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, D.C.: The National Academies Press. doi.org/10.17226/10490).<br>* Other names: Trade name: Glycine (Manufacturer Fagron Inc);|
| Experimental: Alanine (Trade name: L-Alanine)<br>Dietary Supplement: Alanine, water soluble powder, 8.5 g daily in one dose, during 30 days. | Dietary Supplement: Alanine<br>* Amino acids are the building blocks of proteins, they are essential organic compounds in the normal diet, and are also individually administered as nutritional supplements. Alanine will be supplied in the form of water soluble powder (the commercial form), for one daily dose. The corresponding amino acid doses were established based on the highest intakes at the 99th percentile in adult men (Institute of Medicine. 2005. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, D.C.: The National Academies Press. doi.org/10.17226/10490).<br>* Other names: Trade name: L-Alanine (Manufacturer Fagron Inc);|
| Experimental: Leucine (Trade name: L-Leucine)<br>Dietary Supplement: Leucine, water soluble powder, 14 g daily in one dose, during 30 days. | Dietary Supplement: Leucine<br>* Amino acids are the building blocks of proteins, they are essential organic compounds in the normal diet, and are also individually administered as nutritional supplements. Leucine will be supplied in the form of water soluble powder (the commercial form), for one daily dose. The corresponding amino acid doses were established based on the highest intakes at the 99th percentile in adult men (Institute of Medicine. 2005. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, D.C.: The National Academies Press. doi.org/10.17226/10490).<br>* Other names: Trade name: L-Leucine (Manufacturer Fagron Inc);|
| Experimental: Isoleucine (Trade name: L-Isoleucine)<br>Dietary Supplement: Isoleucine, water soluble powder, 8.2 g daily in one dose, during 30 days. | Dietary Supplement: Isoleucine<br>* Amino acids are the building blocks of proteins, they are essential organic compounds in the normal diet, and are also individually administered as nutritional supplements. Isoleucine will be supplied in the form of water soluble powder (the commercial form), for one daily dose. The corresponding amino acid doses were established based on the highest intakes at the 99th percentile in adult men (Institute of Medicine. 2005. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, D.C.: The National Academies Press. doi.org/10.17226/10490).<br>* Other names: Trade name: L-Isoleucine (Manufacturer Fagron Inc);|
| Experimental: Valine (Trade name: L-Valine)<br>Dietary Supplement: Valine, water soluble powder, 9 g daily in one dose, during 30 days. | Dietary Supplement: Valine<br>* Amino acids are the building blocks of proteins, they are essential organic compounds in the normal diet, and are also individually administered as nutritional supplements. Valine will be supplied in the form of water soluble powder (the commercial form), for one daily dose. The corresponding amino acid doses were established based on the highest intakes at the 99th percentile in adult men (Institute of Medicine. 2005. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, D.C.: The National Academies Press. doi.org/10.17226/10490).<br>* Other names: Trade name: L-Valine (Manufacturer Fagron Inc);|
| Experimental: Cysteine (Trade name: L-Cysteine)<br>Dietary Supplement: Cysteine, water soluble powder, 2.2 g daily in one dose, during 30 days. | Dietary Supplement: Cysteine<br>* Amino acids are the building blocks of proteins, they are essential organic compounds in the normal diet, and are also individually administered as nutritional supplements. Cysteine will be supplied in the form of water soluble powder (the commercial form), for one daily dose. The corresponding amino acid doses were established based on the highest intakes at the 99th percentile in adult men (Institute of Medicine. 2005. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, D.C.: The National Academies Press. doi.org/10.17226/10490).<br>* Other names: Trade name: L-Cysteine (Manufacturer Fagron Inc);|
| Experimental: Arginine (Trade name: L-Arginine)<br>Dietary Supplement: Arginine, water soluble powder, 10 g daily in one dose, during 30 days. | Dietary Supplement: Arginine<br>* Amino acids are the building blocks of proteins, they are essential organic compounds in the normal diet, and are also individually administered as nutritional supplements. Arginine will be supplied in the form of water soluble powder (the commercial form), for one daily dose. The corresponding amino acid doses were established based on the highest intakes at the 99th percentile in adult men (Institute of Medicine. 2005. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, D.C.: The National Academies Press. doi.org/10.17226/10490).<br>* Other names: Trade name: L-Arginine (Manufacturer Fagron Inc);|
| Experimental: Methionine (Trade name: DL-Methionine)<br>Dietary Supplement: Methionine, water soluble powder, 4 g daily in one dose, during 30 days. | Dietary Supplement: Methionine<br>* Amino acids are the building blocks of proteins, they are essential organic compounds in the normal diet, and are also individually administered as nutritional supplements. Methionine will be supplied in the form of water soluble powder (the commercial form), for one daily dose. The corresponding amino acid doses were established based on the highest intakes at the 99th percentile in adult men (Institute of Medicine. 2005. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, D.C.: The National Academies Press. doi.org/10.17226/10490).<br>* Other names: Trade name: DL-Methionine (Manufacturer Fagron Inc);|
| Experimental: Glutamate (Trade name: L-Glutamic acid)<br>Dietary Supplement: Glutamate, water soluble powder, 33 g daily in one dose, during 30 days. | Dietary Supplement: Glutamate<br>* Amino acids are the building blocks of proteins, they are essential organic compounds in the normal diet, and are also individually administered as nutritional supplements. Glutamate will be supplied in the form of water soluble powder (the commercial form), for one daily dose. The corresponding amino acid doses were established based on the highest intakes at the 99th percentile in adult men (Institute of Medicine. 2005. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, D.C.: The National Academies Press. doi.org/10.17226/10490).<br>* Other names: Trade name: L-Glutamic acid (Manufacturer Fagron Inc);|
| Experimental: Glutamine (Trade name: L-Glutamine)<br>Dietary Supplement: Glutamine, water soluble powder, 30 g daily in one dose, during 30 days. | Dietary Supplement: Glutamine<br>* Amino acids are the building blocks of proteins, they are essential organic compounds in the normal diet, and are also individually administered as nutritional supplements. Glutamine will be supplied in the form of water soluble powder (the commercial form), for one daily dose. The corresponding amino acid doses were established based on the highest intakes at the 99th percentile in adult men (Institute of Medicine. 2005. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. Washington, D.C.: The National Academies Press. doi.org/10.17226/10490).<br>* Other names: Trade name: L-Glutamine (Manufacturer Fagron Inc);|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Serum atherogenicity | Macrophage lipids (triglycerides and cholesterol) content (μg/mg cell protein) following incubation with serum derived from the subjects. | 1 year | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Serum triglyceride concentration | Triglyceride concentration (mg/dL) vs control group | 1 year | 
| Macrophage cholesterol content | Cholesterol concentration (mg/dL) vs control group | 1 year | 
| Serum oxidation | Serum levels of thiobarbituric acid reactive substances (TBARS) | 1 year | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Amino acids, Macrophage atherogenicity, Triglycerides, Cholesterol, Lipoproteins