Sperm DNA fragmentation and apoptosis in the sperm of men with oligozoospermia are closely related to anti-ODF2 autoantibodies

Amirhossein Kiani; Yusuf Döğüş; Sahar Saadatnia; Yalda Yazdani; Fatemeh Asadi; Bashar Zuhair Talib Al-Naqeeb; Sahand Saeidpour Masouleh; Muna S Merza; Amin Daemi; Asiye Rahimi

doi:10.1016/j.prp.2023.154434

Sperm DNA fragmentation and apoptosis in the sperm of men with oligozoospermia are closely related to anti-ODF2 autoantibodies

Pathol Res Pract. 2023 May:245:154434. doi: 10.1016/j.prp.2023.154434. Epub 2023 Apr 1.

Authors

Affiliations

¹ Department of Immunology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
² Department of Medical Biochemistry, Faculty of Medicine, Cukurova University, Adana, Turkey.
³ Faculty of Nursing and Midwifery, Hamadan University of Medical Sciences, Hamadan, Iran.
⁴ Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
⁵ Department of Genetics, Izeh Branch, Islamic Azad University, Izeh, Iran.
⁶ Anesthesia Technology Department, Al-Turath University College, Al Mansour, Baghdad, Iraq.
⁷ Department of Medical Biotechnologies, University of Siena, 53100 Siena, Italy.
⁸ Prosthetic dental Techniques Departmen, Al-Mustaqbal University College, Babylon, 51001,Iraq.
⁹ Department of Medical Biochemistry, Faculty of Medicine, Cukurova University, Adana, Turkey. Electronic address: phd_bio@yahoo.com.
¹⁰ Midwifery Undergraduate Student, Faculty of Nursing and Midwifery, Mashhad University of Medical Sciences, Mashhad, Iran. Electronic address: aserahimi7@gmail.com.

PMID: 37086632
DOI: 10.1016/j.prp.2023.154434

Abstract

Background: Around 15% of couples of childbearing age suffer from infertility; in 50% of these cases, the male factor is present. In this study, we investigated the association between anti-ODF2 autoantibody existence and the DNA fragmentation and apoptosis of sperm in oligozoospermia men.

Material and methods: 35 fertile men and 57 oligozoospermia men are enrolled in this study as control and case groups, respectively. After the identification of ODF2 as a possible target of anti-sperm antibodies in sera of oligozoospermia men using two-dimensional gel electrophoresis followed by western blotting and mass spectrometry, the case group serums were screened for anti-ODF2 autoantibodies and divided into anti-ODF2 negative (N = 24) and positive (N = 33) subgroups to follow assays. The mRNA expression levels of ODF2, Caspases 3, 8, 9, BAX, and BCL-2 were evaluated via qRT-PCR in spermatozoa samples of mentioned groups. DNA fragmentation and apoptosis rate of spermatozoa in studied groups were assessed using an SDF kit and flow cytometry, respectively.

Results: Mass spectrometry showed that ODF2 is one of the anti-sperm antibodies targeted in oligozoospermia patients. 33 of 57 oligozoospermia men had anti-ODF2 autoantibody in their sera. An elevated expression of ODF2 mRNA was observed in spermatozoa of anti-ODF2⁺ patients compared to anti-ODF2^- patients and controls. There was an increased expression level of Caspase 3, 8, 9, and BAX and decreased expression of BCL-2 in spermatozoa of anti-ODF2⁺ patients compared to anti-ODF2^- patients and controls. Noticeable increases in DNA fragmentation and apoptosis rate of anti-ODF2⁺ patients' spermatozoa were observed compared to anti-ODF2^- patients and healthy controls spermatozoa. A positive correlation was observed between ODF-2 expression and DNF fragmentation and apoptosis rate of anti-ODF2⁺ patients' spermatozoa.

Conclusion: Our results revealed that ODF2 is one of the main spermatozoa structural proteins, which is one of the anti-sperm antibodies targets, and its dysregulated expression may result in an increased rate of sperm DNA fragmentation and apoptosis.

Keywords: 2DE; Apoptosis; DNA fragmentation; ODF2; Oligozoospermia.

MeSH terms

Apoptosis / genetics
Autoantibodies
DNA Fragmentation
Humans
Male
Oligospermia* / genetics
RNA, Messenger
Spermatozoa
bcl-2-Associated X Protein

Substances

Autoantibodies
bcl-2-Associated X Protein
RNA, Messenger
ODF2 protein, human