The Exercise-Induced Poise of Thiol/Disulfide Form of Gluthatione Quantitation by High-Performance Liquid Chromatography with Fluorescence Detection

Document Type : Original research papers

Authors

1 Department of Sports Physiology, Faculty of Educational Sciences and Psychology, University of Mohaghegh Ardabili, Ardabil, Iran

2 Department of Physiology, Faculty of Physical Education and Sport Science, University of Guilan, Rasht, Iran

3 Department of Physiology, Faculty of Medicine, Guilan University of Medical sciences, Rasht, Iran

4 Department of Exercise Physiology, Faculty of Education and Psychology, Mohaghegh Ardabili University, Ardabil, Iran

Abstract

Introduction: Despite beneficial effects of exercise,balance between oxidants and antioxidants have great health importance.Therefore,reduction potentials for redox couples might be useful indicators of health;however exercise-induced poise of thiol/disulfide form gluthatione has not been measured by HPLC. Objective:The aim of this study was quantitation of poise of thiol/disulfide form gluthatione by in vivo Eh values for GSH/GSSG in erythrocytes in subjects with different physical training status.methods:Thirty male subjects participated in this cross-sectional study and were assigned as professional athletes (PA),recreational athletes (RA)and nonathletes (NA groups.Based on self-reported frequency of physical training, subjects from PA group were selected from professional soccer players; subjects from RA group were identified: moderately trained subjects with regular physical training and nonatletes had no physical activity in their routine. Blood samples were taken from an antecubital vein and analyzed into two part: The part 1 were used for measuring hemoglobin and hematocrit using automated Coulter Counter (Sysmex k-x21) and part 2 containing EDTA were centrifuged at 1600 g for 5 min to obtain Erythrocytes, they were washed twice with cold 9% NaCl solution and Erythrocytes lysed by freezing for 2 hours. Finally, hemolysate (100 mL) was deproteinized and used for measurement of oxidized glutathione (GSSG) to reduced glutathione (GSH) by HPLC; The HPLC analyses were performed with Agilent 1200 series HPLC systems equipped with a quaternary pump system (G1311A) and a fluorescence detector (G1321A) (Agilent Technologies, Waldbronn, Germany). Fluorimetric detection was performed at 420 nm after excitation at 340 nm. Then, Eh for GSH/2GSSG was calculated by Nernst equation (Eh=E0 + RT/nF ln([GSSG]/[GSH]2). Results: Recreational athletes had the less amounts of reduction potentials for GSH in compared to nonathletes group; however recreational athletes had the redox environment with the most negative amount of Eh in their erythrocytes. This study suggests that physical activity of individuals determines the reduction potentials for GSH in erythrocytes. Discussion and Conclusion: Long term regular exercise training with moderate intensity has the least reduction potentials that ensuring healthiness, versus intensive exercise training same as passive lifestyle leads to most amounts of reduction potential and may be consequent development of related diseases

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References

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History

  • Receive Date: 06 March 2024
  • Revise Date: 24 March 2026
  • Accept Date: 10 February 2025

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