Association of NOX5 Expression with Sperm Activity and Motility in Pathospermic Infertile Men

Daylan, Benay; Korkmaz, Oya; Tok , Olgu E.; Cakici, Cagri; Yigitbasi, Turkan; Karahasanoglu, Ayse; Yelke, Tuba V.; Ustabas, Tugba S.; Ayla, Sule

Association of NOX5 Expression with Sperm Activity and Motility in Pathospermic Infertile Men

Vol. 25, Issue 3, / July-September 2024
(Original Article, pages 184-192)

Benay Daylan: 1- Department of Histology and Embryology, Istanbul Medipol University, School of Medicine, Istanbul, Turkey; 2- Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, Turkey
Oya Korkmaz: - Department of Histology and Embryology, Malatya Turgut Ozal University, School of Medicine, Malatya, Turkey
Olgu Enis Tok: 1- Department of Histology and Embryology, Istanbul Medipol University, School of Medicine, Istanbul, Turkey; 2- Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, Turkey
Cagri Cakici: 1- Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, Turkey; 2- Department of Biochemistry, Istanbul Medipol University, School of Medicine, Istanbul, Turkey
Turkan Yigitbasi: 1- Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, Turkey; 2- Department of Biochemistry, Istanbul Medipol University, School of Medicine, Istanbul, Turkey
Ayse Karahasanoglu: - Medipol Mega University Hospital, IVF Center, Istanbul, Turkey
Tuba Varlı Yelke: - Medipol Mega University Hospital, IVF Center, Istanbul, Turkey
Tugba Senel Ustabas: - Medipol Mega University Hospital, IVF Center, Istanbul, Turkey
Sule Ayla: - Department of Histology and Embryology, Istanbul Medeniyet University, School of Medicine, Istanbul, Turkey

Received: 2/11/2024 Accepted: 6/29/2024 - Publisher : Avicenna Research Institute

Other Format

Abstract

Background: The newest NOX isoform, NOX5, has been found in mammalian spermatozoa. Many physiological and pathological situations in spermatozoa are mediated by reactive oxygen species (ROS). NOX5 is the main source of ROS in spermatozoa. Our purpose was to investigate the changes in NOX5 expression and the effect of NOX5 expression on sperm motility, chromatin integrity, and oxidative status in oligoasthenozoospermic compared to normozoospermic men.
Methods: Semen samples were collected from 30 normozoospermic (NS) and 30 oligoasthenozoospermic (OAS) men. NOX5 protein expression in sperm samples was evaluated by immunohistochemistry and western blot. Oxidative stress status was evaluated by total antioxidant capacity (TAC), total oxidant capacity (TOC), and oxidative stress index (OSI) parameters. Chromatin integrity in spermatozoa was evaluated by toluidine blue staining.
Results: NOX5 expression levels were significantly higher in OAS group than in NS group (p<0.001). In addition, chromatin integrity was significantly higher in the OAS group in comparison to NS group (p<0.001). TAC levels were higher in the NS group, but OSI and TOC levels were significantly higher in OAS group (p<0.001). It was found that NOX5 protein expression was positively correlated with oxidative stress and chromatin integrity and negatively correlated with motility (p<0.01).
Conclusion: These results suggest that overexpression of NOX5 may be the source of excessive ROS production and oxidative stress injuries in oligoasthenozoospermic men. Considering that NOX5 expression is positively correlated with oxidative stress and chromatin integrity but negatively correlated with motility, it can be considered a biomarker to be used in assisted reproductive procedures.

Keywords: Male infertility, NADPH oxidase 5, Reactive oxygen species, Sperm motility

To cite this article:

Daylan B, Korkmaz O, Tok  OE, Cakici C, Yigitbasi T, Karahasanoglu A, et al. Association of NOX5 Expression with Sperm Activity and Motility in Pathospermic Infertile Men. J Reprod Infertil. 2024;25(3):184-192.

References

Zegers-Hochschild F, Adamson GD, Dyer S, Racowsky C, de Mouzon J, Sokol R, et al. The international glossary on infertility and fertility care. Hum Reprod. 2017;32(9):1786-801. [PubMed]
Hotaling JM. Genetics of male infertility. Urol Clin North Am. 2014;41(1):1-17. [PubMed]
Virtanen HE, Toppari J. Cryptorchidism and fertility. Endocrinol Metab Clin North Am. 2015;44(4):751-60. [PubMed]
Parvin A, Erabi G, Alemi A, Rezanezhad A, Maleksabet A, Sadeghpour S, et al. Seminal plasma proteomics as putative biomarkers for male infertility diagnosis. Clin Chim Acta. 2024;561:119757. [PubMed]
Andrabi SW, Ara A, Saharan A, Jaffar M, Gugnani N, Esteves SC. Sperm DNA fragmentation: causes, evaluation and management in male infertility. JBRA Assist Reprod. 2024;28(2):306-19. [PubMed]
de Lamirande E, Gagnon C. Reactive oxygen species and human spermatozoa. I. effects on the motility of intact spermatozoa and on sperm axonemes. J Androl. 1992;13(5):368-78. [PubMed]
Aitken RJ. Possible redox regulation of sperm motility activation. J Androl. 2000;21(4):491-6. [PubMed]
Griveau JF, Le Lannou D. Effects of antioxidants on human sperm preparation techniques. Int J Androl. 1994;17(5):225-31. [PubMed]
de Lamirande E, Gagnon C. Impact of reactive oxygen species on spermatozoa: a balancing act between beneficial and detrimental effects. Hum Reprod. 1995;10 Suppl 1:15-21. [PubMed]
O'Flaherty C. Redox regulation of mammalian sperm capacitation. Asian J Androl. 2015;17(4):583-90. [PubMed]
Aitken RJ. The role of free oxygen radicals and sperm function. Int J Androl. 1989;12(2):95-7. [PubMed]
Iwasaki A, Gagnon C. Formation of reactive oxygen species in spermatozoa of infertile patients. Fertil Steril. 1992;57(2):409-16. [PubMed]
Bánfi B, Molnár G, Maturana A, Steger K, Hegedûs B, Demaurex N, et al. A Ca(2+)-activated NADPH oxidase in testis, spleen, and lymph nodes. J Biol Chem. 2001;276(40):37594-601. [PubMed]
Setyawan EMN, Kim MJ, Oh HJ, Kim GA, Jo YK, Lee SH, et al. Spermine reduces reactive oxygen species levels and decreases cryocapacitation in canine sperm cryopreservation. Biochem Biophys Res Commun. 2016;479(4):927-32. [PubMed]
Sabeur K, Ball BA. Characterization of NADPH oxidase 5 in equine testis and spermatozoa. Reproduction. 2007;134(2):263-70. [PubMed]
Cheng G, Cao Z, Xu X, van Meir EG, Lambeth JD. Homologs of gp91phox: cloning and tissue expression of Nox3, Nox4, and Nox5. Gene. 2001; 269(1-2):131-40. [PubMed]
BelAiba RS, Djordjevic T, Petry A, Diemer K, Bonello S, Banfi B, et al. NOX5 variants are functionally active in endothelial cells. Free Radic Biol Med. 2007;42(4):446-59. [PubMed]
Brar SS, Corbin Z, Kennedy TP, Hemendinger R, Thornton L, Bommarius B, et al. NOX5 NAD(P)H oxidase regulates growth and apoptosis in DU 145 prostate cancer cells. Am J Physiol Cell Physiol. 2003;285(2):C353-69. [PubMed]
Guzik TJ, Chen W, Gongora MC, Guzik B, Lob HE, Mangalat D, et al. Calcium-dependent NOX5 nicotinamide adenine dinucleotide phosphate oxidase contributes to vascular oxidative stress in human coronary artery disease. J Am Coll Cardiol. 2008;52(22):1803-9. [PubMed]
Peng JJ, Liu B, Xu JY, Peng J, Luo XJ. NADPH oxidase: its potential role in promotion of pulmonary arterial hypertension. Naunyn Schmiedebergs Arch Pharmacol. 2017;390(4):331-8. [PubMed]
Tarafdar A, Pula G. The role of NADPH oxidases and oxidative stress in neurodegenerative disorders. Int J Mol Sci. 2018;19(12):3824. [PubMed]
Wright C, Milne S, Leeson H. Sperm DNA damage caused by oxidative stress: modifiable clinical, lifestyle and nutritional factors in male infertility. Reprod Biomed Online. 2014;28(6):684-703. [PubMed]
Vatannejad A, Tavilani H, Sadeghi MR, Amanpour S, Shapourizadeh S, Doosti M. Evaluation of ROS-TAC Score and DNA damage in fertile normozoospermic and infertile asthenozoospermic males. Urol J. 2017;14(1):2973-8. [PubMed]
World health organization. WHO laboratory manual for the examination and processing of human semen. 6th ed. Geneva: World health organization; 2021. 276 p.
Erel O. A novel automated method to measure total antioxidant response against potent free radical reactions. Clin Biochem. 2004;37(2):112-9. [PubMed]
Erel O. A new automated colorimetric method for measuring total oxidant status. Clin Biochem. 2005;38(12):1103-11. [PubMed]
Kim HS, Kang MJ, Kim SA, Oh SK, Kim H, Ku SY, et al. The utility of sperm DNA damage assay using toluidine blue and aniline blue staining in routine semen analysis. Clin Exp Reprod Med. 2013;40(1):23-8. [PubMed]
Ghani E, Keshtgar S, Habibagahi M, Ghannadi A, Kazeroni M. Expression of NOX5 in human teratozoospermia compared to normozoospermia. Andrologia. 2013;45(5):351-6. [PubMed]
Musset B, Clark RA, DeCoursey TE, Petheo GL, Geiszt M, Chen Y, et al. NOX5 in human spermatozoa: expression, function, and regulation. J Biol Chem. 2012;287(12):9376-88. [PubMed]
Vatannejad A, Tavilani H, Sadeghi MR, Karimi M, Lakpour N, Amanpour S, et al. Evaluation of the NOX5 protein expression and oxidative stress in sperm from asthenozoospermic men compared to normozoospermic men. J Endocrinol Invest. 2019;42(10):1181-9. [PubMed]
Keshtgar S, Ghani E. Impact of calcium and reactive oxygen species on human sperm function: Role of NOX5. Andrologia. 2022;54(8):e14470. [PubMed]
Aitken RJ, Clarkson JS. Significance of reactive oxygen species and antioxidants in defining the efficacy of sperm preparation techniques. J Androl. 1988;9(6):367-76. [PubMed]
Aitken RJ, Clarkson JS, Fishel S. Generation of reactive oxygen species, lipid peroxidation, and human sperm function. Biol Reprod. 1989;41(1):183-97. [PubMed]
De Iuliis GN, Wingate JK, Koppers AJ, McLaughlin EA, Aitken RJ. Definitive evidence for the nonmitochondrial production of superoxideanion by human spermatozoa. J Clin Endocrinol Metab. 2006;91(5):1968-75. [PubMed]
Aitken RJ, Ryan AL, Baker MA, McLaughlin EA. Redox activity associated with the maturation and capacitation of mammalian spermatozoa. Free Radic Biol Med. 2004;36(8):994-1010. [PubMed]
Hong J, Behar J, Wands J, Resnick M, Wang LJ, Delellis RA, et al. Bile acid reflux contributes to development of esophageal adenocarcinoma via activation of phosphatidylinositol-specific phospholipase Cgamma2 and NADPH oxidase NOX5-S. Cancer Res. 2010;70(3):1247-55. [PubMed]
Pourmasumi S, Khoradmehr A, Rahiminia T, Sabeti P, Talebi AR, Ghasemzadeh J. Evaluation of sperm chromatin integrity using aniline blue and toluidine blue staining in infertile and normozoospermic men. J Reprod Infertil. 2019;20(2):95-101. [PubMed]