Thymoquinone improves motility, plasma membrane integrity and DNA integrity of frozen–thawed ram semen

Abstract

The aim of this study was to determine the effects of thymoquinone (TQ), which is the most essential active compound of Nigella sativa, on the spermatological parameters of ram semen during cryopreservation. Ejaculates were collected from five Sonmez rams using an artificial vagina and extended with Tris‐based extender not containing TQ (control, 0 μg/ml TQ) and containing 10, 25, 50 and 100 μg/ml TQ. The extended semen samples were equilibrated in a + 4°C cold cabinet for 2 h. After 2 h, the samples were loaded into 0.25 ml French straws. The straws were frozen by liquid nitrogen vapour and stored in a liquid nitrogen container (−196°C). The frozen straws were thawed in a water bath (37°C for 30 s) and evaluated in terms of motility characteristics, plasma membrane and acrosome integrity, mitochondrial reactive oxygen species levels, lipid peroxidation levels, DNA damage and biochemical alterations (oxidative stress index, malondialdehyde and glutathione). TQ100 had higher total motility (53.59 ± 3.01) and progressive motility (19.84 ± 1.44; not significantly different from TQ25 and TQ50) compared to the control and TQ10 (p ˂ 0.05). According to the results of the analyses on motility characteristics, there were significant differences between the groups in terms of curvilinear velocity (VCL), amplitude of lateral head displacement (ALH) and linearity (LIN; p ˂ 0.05). The highest DNA damage was detected in the control group (p ˂ 0.05). TQ50 had higher plasma membrane and acrosome integrity (59.56 ± 5.92) compared to the control and TQ25 (p < 0.05) but not significantly different from TQ10 and TQ100. The lowest mitochondrial reactive oxygen species levels were detected in TQ50 and TQ100 (p ˂ 0.05). There were no significant differences among the groups in terms of their oxidative stress index, lipid peroxidation, malondialdehyde and glutathione levels (p > 0.05). According to the results, it could be concluded that supplementing 50 or 100 μg/ml TQ to Tris extenders that were used for ram semen cryopreservation showed a positive effect on motility, plasma membrane integrity and acrosome integrity, and it reduced DNA damage and mitochondrial reactive oxygen species levels.

Investigation of Changes in Spermatozoon Characteristics, Chromatin Structure, and Antioxidant/Oxidant Parameters after Freeze-Thawing of Hesperidin (Vitamin P) Doses Added to Ram Semen

“…Vijaya Bharathi et al [47] explained that glucosyl H, a modified form of H, had a protective effect against chromatin damage in the sperm cells of rats, in which reproductive toxicity was induced by increasing cellular antioxidant levels. In accordance with this study, 50 and 100 µg/mL thymoquinone added to Tris semen extender reduced chromatin damage in Sönmez rams in a previous study by our research group [48]. Additionally, the ability of H to stabilize the mitochondrial membrane was reported to protect sperm cells from apoptosis and increase mitochondrial functions [28].…”

Evaluation of the in vitro cryopreservative performance of Juniper berry oil (Juniperus communis) on frozen-thawed bull semen

“…Supporting the DNA integrity result identified in the JB25 and JB50 groups in this study, it was demonstrated that 2, 4 and 6 μg/ml spirulina extract had a protective effect on spermatozoon DNA integrity, and a toxic effect when > 6 μg/ml of it was used (Mizera et al 2019). The findings of this study were in favour of the findings of the studies conducted by our researcher group in recent years, where we have used various antioxidants (rosmarinic acid, pine bark tree extract, vaccenic acid, hesperidin, thymoquinone) due to their remedial effects on DNA integrity (Yeni et al 2018;Taşdemir et al 2020;Avdatek et al 2022;İnanç et al 2022;Yeni et al 2022). It was concluded that as an antioxidant, JB at appropriate doses (25 and 50 μg/ml) maintains the integrity of DNA and this antioxidant property of JB manifests its healing effect on DNA damage by creating an additive effect along with other cryoprotectants in the extender.…”

Effects of sinapic acid on lead acetate‐induced oxidative stress, apoptosis and inflammation in testicular tissue

“…Motility is one of the main parameters used to determine sperm quality. 92 Previous studies report that PbAc reduces sperm motility in rats. 24,90,93 In our study, after PbAc administration, it was determined that sperm total motility decreased significantly, and the percentage of dead and abnormal sperm increased significantly compared to other groups.…”

“…It is thought that this condition originates from the affected seminiferous tubules in the histologically deteriorated testicular tissue. Motility is one of the main parameters used to determine sperm quality 92 . Previous studies report that PbAc reduces sperm motility in rats 24,90,93 .…”