Evaluation of the NOX5 protein expression and oxidative stress in sperm from asthenozoospermic men compared to normozoospermic men

  • A. Vatannejad
  • H. Tavilani
  • M. R. Sadeghi
  • M. Karimi
  • N. Lakpour
  • S. Amanpour
  • M. Shabani Nashtaei
  • M. DoostiEmail author
Original Article



NADPH oxidase 5 (NOX5), the main isoform of NOX in spermatozoa, has been recognized as the main active generators of reactive oxygen species (ROS), including superoxide anion (O 2 −. ) and hydrogen peroxide (H2O2). ROS have been shown to play important roles in many physiological and pathological conditions in spermatozoa. The present study aims to investigate the alterations of NOX5 protein expression and oxidative stress (OS) status in asthenozoospermic men compared to normozoospermic men.


Semen samples were collected from 25 asthenozoospermic men and 28 normozoospermic men. In this study, NOX5 protein expression was evaluated by Western blotting. An OS status was evaluated by measuring of ROS (O 2 −. and H2O2), DNA damage and plasma membrane integrity in spermatozoa.


The protein expression of NOX5 (p < 0.0001) was remarkably higher in asthenozoospermic men in comparison to normozoospermic men. In addition, the percentages of intracellular O 2 −. (p < 0.0001), H2O2 (p < 0.0001) in viable spermatozoa, apoptotic sperm cells with altered plasma membrane (p < 0.001) and DNA damage (p = 0.001) were significantly increased in asthenozoospermic men compared to normozoospermic men.


The present study provides evidence that the overexpression of NOX5 protein may induce excessive ROS production and oxidative stress damages to DNA and plasma membrane integrity in asthenozoospermic men.


NADPH oxidase 5 Male infertility Oxidative stress Reactive oxygen species 



NADPH oxidase


Reactive oxygen species


Superoxide anion


Hydrogen peroxide


Superoxide dismutase


Oxidative stress


Sperm chromatin dispersion


Sperm DNA fragmentation


World Health Organization


2′, 7′-dichlorodihydrofluorescin diacetate




Propidium iodide



We thank Mehdi Baratchian and Fatima Zahraa Fouani for grammatical editing of the manuscript. The authors would like to thank Omid Clinic particularly Dr. Saeidi for providing samples.

Author contribution

AV performed experiments and wrote the manuscript. HT, MD and MRS, contributed equally to this work by helping to design experiments. SA and NL supplied semen samples. MK analyzed data. All authors read and approved the final version of the manuscript.


This work was supported by the Tehran University of Medical Sciences, Tehran, Iran (Grant numbers 21660 and 30756).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Ethical approval

All procedures performed in this study were in accordance with the ethical standards of the ethical committee (no.Ir.tums.rec.1395.2706) of Tehran University of Medical Sciences (Tehran, Iran).

Informed consent

All participants were informed of the study, and a written consent was obtained from them.

Consent for publication

All authors approve the manuscript for publication.


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Copyright information

© Italian Society of Endocrinology (SIE) 2019

Authors and Affiliations

  • A. Vatannejad
    • 1
    • 2
  • H. Tavilani
    • 3
  • M. R. Sadeghi
    • 4
  • M. Karimi
    • 5
  • N. Lakpour
    • 6
    • 7
  • S. Amanpour
    • 8
  • M. Shabani Nashtaei
    • 9
  • M. Doosti
    • 10
    Email author
  1. 1.Department of Comparative Biosciences, Faculty of Veterinary MedicineUniversity of TehranTehranIran
  2. 2.Student’s Scientific Research CenterTehran University of Medical SciencesTehranIran
  3. 3.Urology and Nephrology Research CenterHamadan University of Medical SciencesHamadanIran
  4. 4.Monoclonal Antibody Research CenterAvicenna Research Institute, ACECRTehranIran
  5. 5.Department of Epidemiology and Biostatistics, School of Public HealthTehran University of Medical SciencesTehranIran
  6. 6.Reproductive Research CenterAvicenna Research Institute, ACECRTehranIran
  7. 7.Department of Pathology, Faculty of Medicine, Oncopathology Research CenterIran University of Medical SciencesTehranIran
  8. 8.Cancer Biology Research CentreTehran University of Medical SciencesTehranIran
  9. 9.Department of Infertility, Shariati HospitalTehran University of Medical Sciences Faculty of MedicineTehranIran
  10. 10.Department of Clinical Biochemistry, Faculty of MedicineTehran University of Medical SciencesTehranIran

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