Abstract
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.
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This study is based on the thesis of Andrea Bonvini, openly available in Biblioteca Digital-USP at https://doi.org/10.11606/T.9.2019.tde-26082019-112844.
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Abbreviations
- AT:
-
Acute treatment
- CAT1:
-
Cationic amino acid transporter
- COX-2:
-
Cyclooxygenase 2
- CT:
-
Chronic treatment
- CTL:
-
Control group
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- DMEM-S:
-
Dulbecco’s modified Eagle’s medium with BCAA supplementation
- DMSO:
-
Dimethyl sulfoxide
- ELISA:
-
Enzyme-linked immunosorbent assay
- FBS:
-
Fetal bovine serum
- IFNs:
-
Type I interferons
- IL:
-
Interleukins
- ILE:
-
Isoleucine group
- iNOS:
-
Inducible nitric oxide synthase
- LEU:
-
Leucine group
- LPS:
-
Lipopolysaccharide
- mTOR:
-
Mammalian target of rapamycin
- MTT:
-
(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium
- NADH:
-
Nicotinamide adenine dinucleotide
- NADPH:
-
Nicotinamide adenine dinucleotide phosphate
- NF-kB:
-
Nuclear factor kappa-B
- NO:
-
Nitric oxide
- PT:
-
Pretreatment
- SIRS:
-
Systemic inflammatory response syndrome
- T2D:
-
Type 2 diabetes
- TCA:
-
Tricarboxylic acid cycle
- TLR4:
-
Toll-like receptor 4
- TNF- α:
-
Tumor necrosis factor-alpha
- TSC1:
-
Tuberous sclerosis complex 1
- VAL:
-
Valine group
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This research was funded by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), grant numbers 2016/04910-0 and 2016/11360-6.
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Conceptualization: Andrea Bonvini and Marcelo Macedo Rogero; methodology: Andrea Bonvini, Leonardo Mendes Bella, Primavera Borelli and Ricardo Ambrosio Fock; software: Andrea Bonvini, Audrey Yule Coqueiro and Leonardo Mendes Bella, formal analysis: Andrea Bonvini and Leonardo Mendes Bella; writing—original draft preparation, Andrea Bonvinii and Marcelo Macedo Rogero; writing—review and editing: Andrea Bonvini, Marcelo Macedo Rogero, Audrey Yule Coqueiro, Raquel Raizel, Leonardo Mendes Bella, Primavera Borelli, Ricardo Ambrosio Fock and Julio Tirapegui; Supervision: Marcelo Macedo Rogero, Primavera Borelli, Ricardo Ambrosio Fock and Julio Tirapegui; funding acquisition: Andrea Bonvini, Audrey Yule Coqueiro and Julio Tirapegui.
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Bonvini, A., Rogero, M.M., Coqueiro, A.Y. et al. Effects of different branched-chain amino acids supplementation protocols on the inflammatory response of LPS-stimulated RAW 264.7 macrophages. Amino Acids 53, 597–607 (2021). https://doi.org/10.1007/s00726-021-02940-w
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DOI: https://doi.org/10.1007/s00726-021-02940-w