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Human Reproduction, Vol. 19, No. 1, 129-138, January 2004
© 2004 European Society of Human Reproduction and Embryology

Relationship between ROS production, apoptosis and DNA denaturation in spermatozoa from patients examined for infertility

Mohamed H. Moustafa1,2, Rakesh K. Sharma1, Julie Thornton3, Edward Mascha3, Mohammed A. Abdel-Hafez2, Anthony J. Thomas1 and Ashok Agarwal1,4

1 Center for Advanced Research in Human Reproduction, Infertility, and Sexual Function, Glickman Urological Institute, and 3 Department of Biostatistics and Epidemiology, The Cleveland Clinic Foundation, Cleveland, OH, USA and 2 Cairo University, Cairo, Egypt

4 To whom correspondence should be addressed at: Center for Advanced Research in Human Reproduction, Infertility, and Sexual Function, Glickman Urological Institute and Department of Obstetrics–Gynecology, Cleveland Clinic Foundation, 9500 Euclid Avenue, Desk A19.1, Cleveland, OH 44195, USA. e-mail: agarwaa{at}ccf.org

BACKGROUND: The aim of this study was to examine the role of apoptosis and reactive oxygen species (ROS) in inducing DNA damage in ejaculated spermatozoa. METHODS: We examined ejaculated spermatozoa from 31 patients examined for infertility and 19 healthy donors for apoptosis, production of ROS and DNA damage using annexin V binding, chemiluminescence assay and sperm chromatin structure assay. RESULTS: The percentage of spermatozoa that underwent apoptosis in the whole ejaculate and mature fraction was higher in the patients than in the donors (P < 0.001 and P = 0.009, respectively). Levels of ROS in the whole ejaculate and immature fraction were higher in the patients than in the donors (P = 0.002 and P = 0.009). Apoptosis was significantly correlated with ROS within patients in the whole ejaculate [r (95% confidence interval) = 0.53 (0.19–0.86)] and in the mature [0.71 (0.39–1.00)] and immature spermatozoa [0.75 (0.45–1.00)]. Only apoptosis and the DNA fragmentation index (DFI) were significantly correlated within patients in the whole ejaculate [0.57 (0.18–0.97)]. CONCLUSIONS: DNA damage may be induced by oxidative assault. Apoptosis may not contribute significantly to the DNA damage.

Key words: apoptosis/DNA damage/reactive oxygen species/spermatozoa


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