Şule Ayla1, Gülden Tunalı2, Bülent E Bilgiç2, Kenan Sofuoğlu3, A Arman Özdemir2, Gamze Tanrıverdi4, Semra Özdemir5, B Cem Soner6, Bahar Öztürk5, Serçin Karahüseyinoğlu7, Esra Güler Aslan8, Ismail Seçkin4. 1. Istanbul Medipol University, School of Medicine, Department of Histology and Embryology, Istanbul, Turkey. Electronic address: sayla@medipol.edu.tr. 2. Zeynep Kamil Gynecology and Maternity Training and Research Hospital, IVF-ET Unit, Istanbul, Turkey. 3. Medistate Hospital, IVF-ET Unit, Istanbul, Turkey. 4. Istanbul University Cerrahpaşa Medical Faculty, Department of Histology and Embryology, Istanbul, Turkey. 5. Istanbul University Cerrahpaşa Medical Faculty, Department of Biophysics, Istanbul, Turkey. 6. Necmettin Erbakan University Medical Faculty, Department of Pharmacology, Konya, Turkey. 7. Koç University, School of Medicine, Department of Histology and Embryology Istanbul, Turkey. 8. Afyon Kocatepe University, School of Medicine, Department of Histology and Embryology, Afyon, Turkey.
Abstract
PURPOSE: Sperm processing (e.g., centrifugation) used in preparation for assisted reproduction can result in excessive generation of reactive oxygen species (ROS) and potential sperm damage. The use of antioxidants during sperm processing has been shown to prevent iatrogenic sperm damage, including DNA damage. In this study, we evaluated the effect of caffeic acid phenethyl ester (CAPE) on oxidative stress mediated sperm dysfunction and DNA damage. METHODS: Semen samples were obtained to liquefy at room temperature. After centrifugation and washing protocols, spermatozoa were incubated in a single step supplemented medium with either of 10, 50 or 100 μmol/L CAPE for 2 hours at 36 °C. After incubation period, MDA levels of seminal plasma were measured. The fragmentation in sperm DNA was detected by light microscopy via use of an aniline blue assay, while ultrastructural morphology was analyzed by transmission electron microscopy. RESULTS: Significant increase has been observed in percent chromatin condensation (assessed by aniline blue staining) and Malondialdehyde (Mmol/L) in oligoasthenoteratozoospermia group before the centrifugation (0.57 ± 0.15). Incubation of samples with 100 μmol/L CAPE after centrifugation resulted in a significantly lower percent chromatin condensation compared to samples incubated without CAPE (0.42 ± 0.12) (P < 0.0033). Incubation of all samples with CAPE (10 μmol/L, 50 μmol/L, 100 μmol/L.) after centrifugation resulted in a significantly lower percentage of Malondialdehyde levels. CONCLUSIONS: The data suggests that preincubation of spermatozoa with the antioxidant CAPE offers protection against oxidative DNA damage in vitro.
PURPOSE: Sperm processing (e.g., centrifugation) used in preparation for assisted reproduction can result in excessive generation of reactive oxygen species (ROS) and potential sperm damage. The use of antioxidants during sperm processing has been shown to prevent iatrogenic sperm damage, including DNA damage. In this study, we evaluated the effect of caffeic acid phenethyl ester (CAPE) on oxidative stress mediated sperm dysfunction and DNA damage. METHODS: Semen samples were obtained to liquefy at room temperature. After centrifugation and washing protocols, spermatozoa were incubated in a single step supplemented medium with either of 10, 50 or 100 μmol/L CAPE for 2 hours at 36 °C. After incubation period, MDA levels of seminal plasma were measured. The fragmentation in sperm DNA was detected by light microscopy via use of an aniline blue assay, while ultrastructural morphology was analyzed by transmission electron microscopy. RESULTS: Significant increase has been observed in percent chromatin condensation (assessed by aniline blue staining) and Malondialdehyde (Mmol/L) in oligoasthenoteratozoospermia group before the centrifugation (0.57 ± 0.15). Incubation of samples with 100 μmol/L CAPE after centrifugation resulted in a significantly lower percent chromatin condensation compared to samples incubated without CAPE (0.42 ± 0.12) (P < 0.0033). Incubation of all samples with CAPE (10 μmol/L, 50 μmol/L, 100 μmol/L.) after centrifugation resulted in a significantly lower percentage of Malondialdehyde levels. CONCLUSIONS: The data suggests that preincubation of spermatozoa with the antioxidant CAPE offers protection against oxidative DNA damage in vitro.