Effects of different branched-chain amino acids supplementation protocols on the inflammatory response of LPS-stimulated RAW 264.7 macrophages

• Published in: Amino acids Vol. 53; no. 4; pp. 597 - 607
• Main Authors: Bonvini, Andrea, Rogero, Marcelo Macedo, Coqueiro, Audrey Yule, Raizel, Raquel, Bella, Leonardo Mendes, Fock, Ricardo Ambrosio, Borelli, Primavera, Tirapegui, Julio
• Format: Journal Article
• Language: English
• Published: Vienna Springer Vienna 01.04.2021; Springer Nature B.V
• Subjects: Amino acids; Analytical Chemistry; Biochemical Engineering; Biochemistry; Biomedical and Life Sciences; Cell culture; Cell viability; Chain branching; Chains; Dietary supplements; Enzyme-linked immunosorbent assay; Inflammation; Inflammatory response; Interleukin 10; Interleukin 6; Isoleucine; Life Sciences; Lipopolysaccharides; Macrophages; Neurobiology; Nutritional status; Original Article; Pretreatment; Prostaglandin E2; Proteomics; Tumor necrosis factor-α; Branched-chain amino acids; Prostaglandins; Cytokines; Nitric oxide; Lipopolysaccharide; Inflammation; Macrophages
• ISSN: 0939-4451; 1438-2199
• DOI: 10.1007/s00726-021-02940-w

Although branched-chain amino acids (BCAA) are commonly used as a strategy to recover nutritional status of critically ill patients, recent findings on their role as immunonutrients have been associated with unfavorable outcomes, especially in obese patients. The present study aimed to explore the effects of different BCAA supplementation protocols in the inflammatory response of LPS-stimulated RAW 264.7 macrophages. Cell cultures were divided into five groups, with and without BCAA supplementation, (2 mmol/L of each amino acid). Then, cell cultures followed three different treatment protocols, consisting of a pretreatment (PT), an acute treatment (AT), and a chronic treatment (CT) with BCAA and LPS stimulation (1 µg/mL). Cell viability was analyzed by MTT assay, NO production was assessed by the Griess reaction and IL-6, IL-10, TNF-α and PGE2 synthesis, was evaluated by ELISA. BCAA significantly increased cell viability in AT and CT protocols, and NO and IL-10 synthesis in all treatment protocols. IL-6 synthesis was only increased in PT and CT protocols. TNF-α and PGE2 synthesis were not altered in any of the protocols and groups. BCAA supplementation was able to increase both pro and anti-inflammatory mediators synthesis by RAW 264.7 macrophages, which was influenced by the protocol applied. Moreover, these parameters were significantly increased by isoleucine supplementation, highlighting a potential research field for future studies.