THE EFFECT OF BCAA SUPPLEMENTATION UPON THE IMMUNE RESPONSE OF TRIATHLETES.

 

Reinaldo Abunasser BASSIT

Mara Assis MALVERDI

Bassit, R.A.2,4; Sawada, L.A.3; Bacurau, R.F.P.1,4; Navarro, F.2,4 & Costa Rosa, L.F.B.P.2,4

1Department of Physiology and Biophysics and 2Department of Histology and Embryology, Institute of Biomedical Sciences, University of Sao Paulo, Brazil.

3Department of Biodynamic of the Movement of the Human Body, School of Sport and Physical Education, University of Sao Paulo, Brazil.

4Laboratory of Human Nutrition for Athletes - CEPEUSP, University of São Paulo, Brazil

Short tittle: BCAA supplementation and immune response

Key words: triathlon, immune system, glutamine, immunossupression, cytokines, lymphocyte proliferation

Correspondence to:

  • Dr. Luís F. B. P. Costa Rosa, Departamento de Histologia e Embriologia, Instituto de Ciências Biomédicas I, Universidade de São Paulo, Av. Lineu Prestes, 1524, 05508-900, Butantã, São Paulo, SP, Brasil.
  • Phone/fax: 55-011-818.7227

Matherial and Methods

Subjects and protocol - The experimental protocol was approved by the local Ethics Committee and after signing an Informed Consent term, twelve elite male triathletes of mean age 25.5 ± 3.2 years (range 21.4-30.1 years) swam 1.5km, cycled 40km and ran 10km (Olympic Triathlon) in the São Paulo International Triathlon held in April 1997 and April 1998.

The athletes were allowed to drink and eat normally, but received branched chain amino acids (BCAA) or placebo for 30 days prior to the competition and one week after the event. BCAA was given twice a day, after each training session (6.0g - 60% L-leucine, 20% L-valine and 20% L-isoleucine) during the first 30 days, and a single dose of 3.0g 30 min before the triathlon, as well as a single dose (3.0g) daily, in the morning, in the first week after the test were administered. On the day of the competition blood samples were collected (20 ml) from the antecubital vein 45 min before the event and 15 min after the race. During the 37 days period the athletes answered a questionnaire reporting their health conditions (Table 1).

Incorporation of [2-14C]-thymidine into peripheral blood lymphocytes - Peripheral blood lymphocytes were cultured in RPMI-1640 medium for 24 h at 37oC in an artificially humidified atmosphere of 5 per cent CO2 in air, under sterile conditions. The cells were cultured in a LAB-LINE Microprocessor CO2 incubator (LAB LINE, U.S.A) in 96 well plates (Corning, NY, U.S.A.), 1 x 105 cells per well (total volume, 200 m l). After 24 h in culture, more than 98 per cent of lymphocytes were viable, as measured by Tripan blue exclusion test.

The cells were pulsed with 20 m l of 0.02 m Ci [2-14C]-thymidine (sp. Act. 56.0 mCi nM-1) diluted in sterile PBS, yelding a final concentration of 1 m g ml –1. Cells were then maintained under these conditions for an additional 15h and harvested automatically by a multiple cell harvester onto a filter paper (cat no 11731 Skatron Combi, Suffolk, U.K.). The paper discs containing the labelled cells were added to vials containing in 5 ml of Bray’s scintillation cocktail, (60 g l-1 naphathalene, 4 g l-1 2,5-diphenyloxazole (PPO), 20 mg l-1 1,4-di-[2-(5-phenyloxazolyl)]-benzene - POPOP, 10 per cent methanol (by vol.) and 2 per cent ethylene glycol (by vol.) in p-dioxan (chromatographic grade) and counted in a Beckman-LS 5000TD liquid scintillator ion counter (Beckman Instruments, Fullerton, CA, U.S.A.). All the reagents used in the preparation of the Brays solution were obtained from Sigma (U.S.A.) or Merck (Darmstadt, Germany).

Measurement of plasmatic glutamine concentration. Plasmatic glutamine concentration was measured enzimaticaly as described by Windmueller & Spaeth (40).

Determination of cytokines concentration. Each 5ml blood sample was transferred to a glass tube containing 5m l of heparin (500 IU/ml). The tubes were kept on ice until centrifuging at 2500 rpm for 8min. The plasma was stored at -80° C. The concentration of cytokines in plasma was measured using commercially available ELISA-kits (Amershan-Life Science): interleukin 1 (IL-1), interleukin 2 (IL-2), g -interferon (IFN) and tumour necrosis factor-a (TNF).

Cytokines produced by cultivated peripheral blood lymphocytes were also measured. Lymphocytes were prepared by centrifuging the blood in the presence of Hystopaque (1.007), for 15 min at 2500 rpm. The mononuclear cells (± 97% lymphocytes) were plated (1.0 x 106 cells/ml) onto a plastic Petri dish in the presence of phytohemaglutinin (PHA) 10 m g/ml to stimulate IL-2, INF and TNF production or lipopolysaccharide (LPS) 10 m g/ml to stimulate IL-1 production. After 48 h, the concentration of the cytokines was measured in the supernatant.

Statistical Analysis - The data obtained in the two events were compared using paired t-test, and the level of significance of p<0.05 was chosen for all statistical comparisons. The data are presented as mean ± SEM.

Table 1. Table of symptoms to be filled by the athletes during 37 days, 30 days prior and one week after an Olympic Triathlon. Each mark corresponds to one point

Symptoms

Days

 

 

 

 

 

 

 

 

 

 

 

 

Fever (oC)

 

 

 

 

 

 

Persistent muscle soreness or tenderness (>than 8 hours)

 

 

 

 

 

 

Pain in the next exercise session

 

 

 

 

 

 

Painful throat

 

 

 

 

 

 

Catarrh in the throat

 

 

 

 

 

 

Itching or sensation of ardour in the throat

 

 

 

 

 

 

Cough

 

 

 

 

 

 

Sneeze

 

 

 

 

 

 

Cephalitis

 

 

 

 

 

 

Coryza

 

 

 

 

 

 

Cold

 

 

 

 

 

 

Grippe

 

 

 

 

 

 

Herpes

 

 

 

 

 

 

Glossitis

 

 

 

 

 

 

Aphtha

 

 

 

 

 

 

Conjunctivitis

 

 

 

 

 

 

Otitis

 

 

 

 

 

 

Mycosis

 

 

 

 

 

 

Infection by Candida

 

 

 

 

 

 

 

 

 

 

 

 

 

Tendinitis

 

 

 

 

 

 

Articular pain

 

 

 

 

 

 

Abrupt change of humour

 

 

 

 

 

 

Insomnia (how many hour without sleep)

 

 

 

 

 

 

Weakness

 

 

 

 

 

 

Anorexia

 

 

 

 

 

 

 

Reinaldo Abunasser BASSIT

 

 

CRN - 6845

Formação Profissional

  • Professor de Educação Física - Universidade de São Paulo - Escola de Educação Física. (EEFUSP) - 1988-1991.
  • Nutricionista - Universidade Anhembi Morumbi - 1993 - 1996
  • Mestrado - Universidade de São Paulo - Instituto de Ciências Biomédicas. (ICB- USP). Laboratório de Metabolismo
    • Ano de entrada: 1998
    • Previsão de Conclusão: 2000

Atividades Profissionais

  • Professor de Natação e Hidroginástica Período: 2º semestre de 1990
  • Preparação Física Individual (Personal Training) Período: 1989 a 1997
  • Professor de Musculação e Condicionamento Físico Período: 1992 a 1995
  • Coordenador de Natação e Hidroginástica Período: 1994
  • Avaliação e Orientação Nutricional dos Atletas Olímpicos
    • Roberto Scheidt
    • Aurélio Miguel
    • Daniele Zangrando
    • Cristiane Parmiggiane
    • Henrique Guimarães
    • Leandro Macedo
    • Mariana Ohata
  • Orientador Nutricional e Avaliador da Composição Corporal de Times Profissionais.
  • Olympikus Telesp (voleibol) 1995 e 1997
  • Sport Clube Banespa (voleibol) 1996 e 1997.
  • Sport Club Corinthians Paulista (Futebol) 1997
  • Portuguesa Futebol Clube 1997.
  • São Paulo Futebol Clube 1995 e 1996.
  • Preparador Físico das Equipes Masculina e Feminina de Handebol Profissional de Osasco. Período: 1997. (Campeão Brasileiro com a Equipe Feminina).
  • Nutricionista da Equipe Brasileira de Duplas na Travessia da Race Across América 98. Período: 1998. (O Brasil foi campeão pela primeira vez obtendo o record na categoria com o tempo de 7 dias e10 horas.)
  • Consultório particular (desde 1997)

 

Mara Assis MALVERDI

CRN - 6844

Formação Profissional

  • Professora de Educação Física - Universidade de São Paulo - Escola de Educação Física. (EEFUSP) - 1988-1991.
  • Nutricionista - Universidade Anhembi Morumbi - 1993 - 1996

Atividades Profissionais

Área de Educação Física

  • Professora de Ed. Física Escolar (1992-1994)
  • Professora de Natação e Hidroginástica (1992-1994)
  • Professora de Ginástica (1992-1994)
  • Coordenadora de atividades aquáticas (1994)

Área de Nutrição

  • Atendimento em clínicas médicas (1997-1998)
  • Atendimento em academias:
    • Master Academia (1997 - 1998)
    • Raia 4 Morumbi (1997-2000)
    • Raia 4 Moema (1997-2000)
    • Kainágua Fitness (desde 1998)
    • Aquademia Sports (1998-1999)
    • Consultório particular (desde 1997)

 

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