RESUMO
This study evaluated the effects of RRR-alpha-tocopherol (500 IU/day, 8 days) on in vivo cytokine response and cytoplasmic expression of inducible nitric oxide synthase (iNOS) and the antioxidant stress protein heme oxygenase-1 (HO-1) in human leukocytes after exhaustive exercise. Thirteen men were investigated in a double-blind, placebo-controlled, cross-over study with a wash-out period of 28 days. The exercise procedure consisted of an incremental treadmill test followed by a continuous run until exhaustion at 110% of the individual anaerobic threshold (total duration 28.5 +/- 0.8 min). HO-1 and iNOS protein were assessed in mono- (M), lympho-, and granulocytes (G) using flow cytometry. Plasma interleukin-6 (IL-6) and IL-8 were measured by ELISA. IL-6 rose significantly whereas IL-8 did not exhibit significant changes after exercise. Changes of IL-6 were not affected by RRR-alpha-tocopherol. Exercise induced an increase of iNOS protein primarily in M and G. A small, but significant, increase of HO-1 protein was measured in M and G. RRR-alpha-Tocopherol did not show any significant effects on cytoplasmic expression of iNOS and HO-1 at rest and after exercise. In conclusion, exhaustive exercise induces expression of iNOS and HO-1 in human leukocytes by a mechanism that is not sensitive to RRR-alpha-tocopherol supplementation.
Assuntos
Antioxidantes/farmacologia , Exercício Físico/fisiologia , Heme Oxigenase (Desciclizante)/biossíntese , Leucócitos/enzimologia , Óxido Nítrico Sintase/biossíntese , Vitamina E/farmacologia , Adulto , Limiar Anaeróbio/efeitos dos fármacos , Antioxidantes/administração & dosagem , Método Duplo-Cego , Indução Enzimática/efeitos dos fármacos , Teste de Esforço , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Granulócitos/efeitos dos fármacos , Granulócitos/enzimologia , Heme Oxigenase (Desciclizante)/genética , Heme Oxigenase-1 , Humanos , Interleucina-6/sangue , Interleucina-8/sangue , Leucócitos/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Linfócitos/enzimologia , Masculino , Proteínas de Membrana , Monócitos/efeitos dos fármacos , Monócitos/enzimologia , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase Tipo II , Corrida , Vitamina E/administração & dosagem , Vitamina E/sangueRESUMO
Previous research revealed an increased expression of HSP72 in leukocytes after vigorous endurance exercise. We questioned whether more intensive but shorter exercise also induces leukocyte HSP72 synthesis. To delineate the role of reactive oxygen species (ROS) in exercise-related HSP72 induction, we additionally examined the effect of RRR-alpha-tocopherol (alpha-toc) on HSP72 expression using a double-blind placebo (P) controlled cross-over design. After supplementation with alpha-toc (500 I.U. daily) or P for 8 days, 9 male subjects performed a combined exhaustive treadmill protocol (total duration 29.4 +/- 2.0 min). HSP72 was assessed on mRNA (RT-PCR) and protein levels (flow cytometry). HSP72 mRNA rose 3 h after exercise only in the P group, but individual differences (alpha-toc - P) did not reveal significant treatment effects. A moderate but significant rise of HSP72 protein occurred in granulocytes up to 48 h after exercise. Three hours post-exercise, granulocyte HSP72 protein was lower when subjects received alpha-toc, but this effect vanished 24 and 48 h post-exercise. Exhaustive treadmill exercise augments HSP72 mRNA in leukocytes and induced a moderate but prolonged response of granulocyte HSP72 protein. These exercise effects are lower when compared to earlier findings obtained after vigorous endurance exercise. ROS seem to be involved, but do not play the major role in the induction of granulocyte HSP72 synthesis after exhaustive exercise.