Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-04T17:06:18Z No. of bitstreams: 2 rosalina_tossige_gomes.pdf: 1360296 bytes, checksum: 20d9d22baa3b186a5cd4139e7af933da (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-04T17:06:52Z (GMT) No. of bitstreams: 2 rosalina_tossige_gomes.pdf: 1360296 bytes, checksum: 20d9d22baa3b186a5cd4139e7af933da (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-01-04T17:06:52Z (GMT). No. of bitstreams: 2 rosalina_tossige_gomes.pdf: 1360296 bytes, checksum: 20d9d22baa3b186a5cd4139e7af933da (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015 / O exerc?cio intervalado de alta intensidade (HIIE) ? caracterizado por breves e repetidas sess?es de exerc?cio intenso, intercaladas por per?odos de repouso ou exerc?cio de baixa intensidade. ? uma modalidade de exerc?cio que tem ganhado muito destaque nos ?ltimos anos por ser uma modalidade de treinamento de baixo volume, embora pouco se saiba a respeito dos seus efeitos na fun??o imune e no estado redox celular. Assim, este estudo avaliou o efeito de uma sess?o de HIIE sobre o estado redox e a fun??o de linf?citos em homens jovens sedent?rios. Esse trabalho foi dividido em dois estudos. No estudo 1 avaliou-se o efeito do HIIE sobre a prolifera??o e produ??o de citocinas e o estado redox de c?lulas mononucleares do sangue perif?rico (PBMC). No estudo 2 avaliou-se o efeito do HIIE sobre a viabilidade e express?o de marcadores de ativa??o em linf?citos. As sess?es de HIIE foram realizadas em bicicleta ergom?trica, e consistiram em oito s?ries de 1 min a 90-100% de pot?ncia pico, com 75 segundos de recupera??o ativa, a 30W, entre as s?ries. O sangue venoso foi colhido antes, imediatamente ap?s e 30 minutos ap?s a sess?o de HIIE. Para avalia??o da prolifera??o celular, por citometria de fluxo, as PBMC foram coradas com Carboxifluoresce?na Succinimidil Ester (CFSE) (10 ?M) e estimuladas com o superant?geno SEB (100 ng/mL), durante 5 dias a 37? C, 5% de CO2. A produ??o de IL-2 e IFN-? em resposta a estimula??o por SEB durante 18 horas, foi avaliada por ELISA. O estado redox celular foi avaliado pela mensura??o da atividade das enzimas catalase (CAT) e super?xido dismutase (SOD), a concentra??o de subst?ncias que reagem ao ?cido tiobarbit?rico (TBARS) e conte?do de glutationa reduzida (GSH). Para avalia??o da viabilidade celular, por citometria de fluxo, as c?lulas foram marcadas com anticorpos anti-Anexina V FITC e iodeto de prop?deo. Para an?lise da ativa??o dos linf?citos, por citometria de fluxo, as PBMC foram estimuladas com SEB por 18 horas, e em seguida marcadas com anticorpos fluorescentes dirigidos contra CD4, CD8, CD19, CD25 e CD69. Os dados foram analisados utitlizando os testes one-way ou two-way ANOVA, considerando p ? 0,05. O HIIE promoveu redu??o na prolifera??o de linf?citos (p = 0,01), aumento na concentra??o de IL-2 (p = 0,02), e desequil?brio redox nas PBMC, marcado por aumento nas concentra??es de TBARS (p = 0,02) e diminui??o na atividade da CAT (p = 0,04). O HIIE n?o alterou a viabilidade das PBMC, mas a frequ?ncia de c?lulas CD4 e CD19, positivas para os marcadores CD25 e CD69, foi menor ap?s o exerc?cio. Contudo, como foi observada redu??o na frequ?ncia de c?lulas CD4+ e CD19+, a redu??o da frequ?ncia de express?o de marcadores de ativa??o refletiu a redu??o do n?mero de c?lulas, e n?o da resposta ao est?mulo superantig?nico. Nossos resultados mostram portanto, que apesar do HIIE promover desequil?brio redox nas PBMC a resposta dos linf?citos ao est?mulo superantig?nico n?o ? alterada. A redu??o da resposta proliferativa ?, provavelmente, reflexo da altera??o da distribui??o das subpopula??es linfocit?rias em decorr?ncia do HIIE. / Disserta??o (Mestrado) ? Programa Multic?ntrico de P?s-gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2015. / ABSTRACT High-intensity interval exercise (HIIE) is characterized by brief and repeated intense exercise sessions, interspersed with periods of rest or low intensity exercise. This exercise modality has gained much attention in recent years, although little is known about its effects on immune function and cellular redox state. This study evaluated the effect of HIIE on the redox state and lymphocyte function in sedentary young males. This work was divided in two studies. The firsty study evaluated the effect of HIIE on peripheral blood mononuclear cells (PBMC) proliferation, cytokine production and redox status. The second study evaluated the effect of HIIE on lymphocyte viability and activation markers expression. HIIE was performed on cycloergometer, and consisted of eight series of 1 min at 90-100% of peak power, with 75 seconds of active recovery, at 30W, between sets. Venous blood was collected before, immediately after and 30 minutes after HIIE. For cell proliferation evaluation by flow cytometry, PBMC were stained with Carboxyfluorescein Succinimidyl Ester (CFSE) (10 mM) and stimulated with the superantigen SEB (100 ng/ml) for 5 days at 37? C, 5% CO2. IL-2 and IFN-? secretion in response to SEB stimulation for 18 hours was assessed by ELISA. The cellular redox status was assessed by measuring the activity of catalase (CAT) and superoxide dismutase (SOD), the concentration of thiobarbituric acid reactive substances (TBARS) and reduced glutathione content (GSH). To assess cellular viability, using flow cytometry, cells were labeled with anti-Annexin V-FITC and propidium iodide. To analyze lymphocyte activation, by flow cytometry, PBMC were stimulated with SEB for 18 hours, and then stained with fluorescent antibodies directed against CD4, CD8, CD19, CD25 and CD69. One-way or two-way ANOVA was employed for statistical analyzis, with ? ? 0.05. Lymphocyte proliferation was reduced (p = 0.01) after HIIE, despite increased IL-2 concentration (p = 0.02), and HIIE also induced PBMC redox imbalance characterized by increased TBARS concentration (p = 0.02) and decreased CAT activity (p = 0.04). PBMC viability was not affected by HIIE, but the frequency of CD4+CD25+/CD69+ and CD19+CD25+/CD69+ cells in response to SEB stimulation was lower after exercise. However, as CD4+ and CD19+ frequencies were reduced, reduced activation markers expression was a consequence of cell number reduction. Our results therefore show that, although HIIE induced redox imbalance, lymphocyte response to superantigen stimulation was not affected. Reduced lymphocyte proliferative response after HIIE is probably due to modifications in lymphocyte subpopulations distribution.