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Effects of antioxidants on post-thawed bovine sperm and oxidative stress parameters: Antioxidants protect DNA integrity against cryodamage
Scopus
Toplam 189 atıf DOI
This study was conducted to determine the effects of methionine, inositol and carnitine on sperm (motility, abnormality, DNA integrity and in vivo fertility) and oxidative stress parameters (lipid peroxidation, total glutathione and antioxidant potential levels) of bovine semen after the freeze-thawing process. Nine ejaculates, collected with the aid of an artificial vagina twice a week from each Simmental bovine, were included in the study. Each ejaculate, splitted into seven equal groups and diluted in Tris-based extender containing methionine (2.5 and 7.5. mM), carnitine (2.5 and 7.5. mM), inositol (2.5 and 7.5. mM) and no additive (control), was cooled to 5 °C and then frozen in 0.25. ml straws. Frozen straws were then thawed individually at 37 °C for 20. s in a water bath for the evaluation. The extender supplemented with 7.5. mM doses of carnitine and inositol led to higher subjective motility percentages (61.9 ± 1.3% and 51.3 ± 1.6%) compared to the other groups. The addition of methionine and carnitine at doses of 2.5 and 7.5. mM and inositol at doses of 7.5. mM provided a greater protective effect in the percentages of total abnormality in comparison to the control and inositol 2.5. mM ( P< 0.001). As regards CASA motility, 7.5. mM carnitine (41.6 ± 2.9% and 54.2 ± 4.9%) and inositol (34.9 ± 2.0% and 47.3 ± 2.2%) caused insignificant increases in CASA and total motility in comparison to the other groups. All of the antioxidants at 2.5 and 7.5. mM resulted in lower sperm with damaged DNA than that of control, thus reducing the DNA damage ( P< 0.05). No significant differences were observed in CASA progressive motility and sperm motion characteristics among the groups. In fertility results based on 59-day non-returns, no significant differences were observed in non-return rates among groups. As regards biochemical parameters, supplementation with antioxidants did not significantly affect LPO and total GSH levels in comparison to the control group ( P> 0.05). The maintenance of AOP level in methionine 2.5. mM was demonstrated to be higher (5.06 ± 0.38. mM) than that of control (0.96 ± 0.29. mM) following the freeze-thawing ( P< 0.001). Supplementation with these antioxidants prior to the cryopreservation process protected the DNA integrity against the cryodamage. Furthermore, future research should focus on the molecular mechanisms of the antioxidative effects of the antioxidants methionine, carnitine and inositol during cryopreservation. © 2010 Elsevier Inc.
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IMMUNOFLUORESCENCE AND BIOCHEMICAL INVESTIGATION OF THE PROTECTIVE EFFECTS OF NARINGIN AND DIOSMIN ON THE FREEZABILITY OF MERINO RAM SEMEN
Scopus
Havuzumuzda 2 atıf almış
BACKGROUND: Various antioxidant substances are added to sperm extenders to protect spermatozoa against oxidative stress and cryodamage. OBJECTIVE: To investigate the effects of the flavonoid diosmin (DIO) and a flavanone glycoside naringin (NAR) on the freezability of ram semen. MATERIALS AND METHODS: In this study, six Merino rams were used during the breeding season. The ejaculates were pooled after collection from the rams. Pooled ejaculates were divided into six groups: control, NAR 1 mM, NAR 2 mM, NAR 4 mM, DIO 2 mM, and DIO 4 mM, and then diluted with a TRIS-based diluent. The pooled semen was equilibrated, placed in 0.25 mL pipettes with 10 × 107 sperm cells in each pipette, and frozen in liquid nitrogen vapor. After 24 h, the pipettes were thawed at 37°C for 25 s and analyzed in terms of spermatological parameters. RESULTS: The highest plasma membrane integrity ratio was found in the DIO 4 mM group, whereas a statistically significant difference was found between the NAR 1 mM and NAR 2 mM groups (p<0.05). While the DIO 4 mM group had the highest acrosome integrity rate, a statistically significant difference was found between the other groups (p<0.05). Mitochondrial activity was the highest in the NAR 4 mM, DIO 4 mM and DIO 2 mM groups (p<0.05). In the analysis of the sperm membrane lipid profile, it was observed that the DIO group had the highest lipid-phospholipid ratio. In sperm membrane protein profile analysis, it was found that both additives exerted protective effects at different levels. The highest total protein content was seen in the DIO 4 mM and NAR 4 mM groups. 8-hydroxydeoxyguanosine (8-OhDG) positivity was more common in the control group than in the DIO and NAR groups. Cu-Zn superoxide dismutase (SOD) expression was lower in the control group and more intense in all other groups. Positive results were especially observed in the acrosome of the sperm cells. CONCLUSION: The addition of NAR and DIO to the ram semen extender increased the quality of sperm parameters after the freeze-thaw process.
Atıf Yapan Makale Bilgileri
Kurumlar (6)
Atatürk Üniversitesi
Erzurum, Turkey
Cumhuriyet Üniversitesi
Sivas, Turkey
Erciyes Üniversitesi
Kayseri, Turkey
Kastamonu University
Kastamonu, Turkey
Provincial Directorate of Agriculture
Mus, Turkey
Selçuk Üniversitesi
Selçuklu, Turkey