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Effects of branched chain amino acids in the inflammatory response induced by lipopolysaccharide (LPS) in a macrophage cell line raw 264.7

Grant number: 16/11360-6
Support Opportunities:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): March 01, 2017
Effective date (End): June 30, 2019
Field of knowledge:Health Sciences - Nutrition - Nutrition Biochemistry
Principal Investigator:Julio Orlando Tirapegui Toledo
Grantee:Andrea Bonvini
Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil


Nutritional therapy is critical to the recovery of ill patients, as it avoids the worsening of the disease and prevents malnutrition. However, when the supply of nutrients, particularly amino acid, is less than ideal, there is a commitment from the individual's immune response to increase susceptibility to infections. Recent studies have discovered the interaction of the amino acids and the immune system, obtaining satisfactory results regarding the modulation promoted by them. However, the literature lacks studies that evaluate the immunomodulatory effects of Branched Chain Amino Acids (BCAA), because, due to their different metabolism, dependent skeletal muscle and not of hepatic oxidation, research is concentrated within the sports nutrition. The exploitation of these possible effects is crucial to the progress of treatment for patients in nutritional therapy since it can provide a significant change in clinical outcomes of sickness, as well as offer an improvement in their quality of life. The aim of this work is to study the effects of branched chain amino acids on the functionality and inflammatory response induced by lipopolysaccharide (LPS) in macrophages cell line RAW 264.7. Cell cultures will be divided into two stages, the first with no LPS and the second with LPS (1¼g / ml) for 30 minutes to evaluate the protein expression in pathway of NF- kB pathway, for 4 hours to gene expression by RT-PCR and 24 hours for analysis of phagocytic and fungicidal activity, cytokine synthesis (TNF-±, IL -6 and IL -10), nitric oxide dosage and cell viability. Each step will be divided into 5 groups: CTL without BCAA supplementation; LEU, supplemented with 1,2mmol/L of leucine; ISO, supplemented with 1,2mmol/L of isoleucine; VAL supplemented with 1,2mmol/L of valine and LIV supplemented with 1,2mmol/L of leucine, 1,2mmol/L of isoleucine and 1,2mmol/L of valine. The cell culture medium (DMEM) has in its normal composition, 0,8mmol/L each of BCAA, so the final concentration in the supplemented groups will be 2 mmol/L. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
BONVINI, ANDREA; COQUEIRO, AUDREY Y.; TIRAPEGUI, JULIO; CALDER, PHILIP C.; ROGERO, MARCELO M.. Immunomodulatory role of branched-chain amino acids. NUTRITION REVIEWS, v. 76, n. 11, p. 840-856, . (16/04910-0, 16/11360-6)
BONVINI, ANDREA; ROGERO, MARCELO MACEDO; COQUEIRO, AUDREY YULE; RAIZEL, RAQUEL; BELLA, LEONARDO MENDES; FOCK, RICARDO AMBROSIO; BORELLI, PRIMAVERA; TIRAPEGUI, JULIO. Effects of different branched-chain amino acids supplementation protocols on the inflammatory response of LPS-stimulated RAW 264.7 macrophages. Amino Acids, v. 53, n. 4, SI, . (16/11360-6, 16/04910-0)

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