Influence of Trolox and Alpha-Lipoic Acid on Post-Thawed Pırlak Ram Sperm Parameters, Oxidative Stress and DNA Damage in Non-Breeding Season

Abstract

The present study was conducted to examine the protective roles of trolox and alpha-lipoic on post-thawed Pirlak ram sperm parameters, oxidative stress and DNA damage in non-breeding season. Semen samples from 5 healty Pırlak rams (2-3 years of age) were used in the study. Six ejaculates for each rams were collected and pooled. Ejaculates were split into three aliquots and diluted to a final concentration of 150x10⁶ spermatozoa/ml with the base extender containing 1 mM alpha lipoic acid, 1 mM trolox and no additive (control) were cooled to 5 °C and equilibrated for 3 h then frozen in 0.25 ml French straws. Samples were thawed in 37 ^oC and evaluated in terms of spermatological parameters, DNA damage, oxidative stress. Sperm motility was increased, acrosome rate and DNA damage were decreased significantly (p<0.05) in ALA, head abnormal sperm rate and membrane integrity were increased, acrosome rate and DNA damege were decreased significantly (P < 0.05) in trolox when compared to the control group. Compared with the control group, reductions in spermatozoon MDA and GSH levels were statistically significant (P <0.05) in lipoic acid and trolox groups. Results of this study suggest that alpha-lipoic acid and trolox improve sperm parmeters, oxidative stress and DNA damage in non-breeding season.

Keywords

• Alpha-lipoic acid,Non-breeding season,Ram sperm,trolox

Sezon Dışı Pırlak Koç Sperma Sulandırıcısına İlave Edilen Lipoik Asid ve Trolox’un Dondurma ve Çözdürme Sonrası Spermatolojik Parametreler, Oksidatif Stres ve DNA Hasarı Üzerine

Abstract

Bu çalışmanın amacı sezon dışı dönemde Pırlak koç sperma sulandırıcısına ilave edilen trolox ve alfa lipoik asitin (ALA) çözüm sonu spermatolojik parametreler, oksidatif stres ve DNA hasarı üzerine etkileri araştırıldı. Çalışmada sperma örnekleri 5 sağlıklı Pırlak koçtan (2-3 yaş) elde edildi ve her koçtan 6 ejakulat toplanarak karıştırıldı. Ejakulatlar ml’de 150x10⁶ spermatozoon olacak şekilde antioksidan içermeyen (kontrol) ve antioksidan içeren 1 mM ALA ve 1 mM trolox sulandırıcılar ile sulandırıldı. Sulandırılan örnekler 0,25 ml’lik payetlerde 5 ⁰C’de 3 saat ekilibrasyona tabi tutularak  sıvı azot buharında donduruldu,37 ^oC'de çözdürülerek spermatolojik parametreler, DNA hasarı, oksidatif stres açısından değerlendirildi. ALA grubunda kontrol grubuna göre spermatozoa motilitesi, akrozom ve DNA bütünlüğü  istatistiki (p<0.05) olarak daha yüksek bulundu. Trolox grubunda kontrol grubuna göre daha yüksek akrozom ve membran bütünlüğü ile  daha az DNA hasarı tespit edildi.  (p<0.05) . Kontrol grubu ile karşılaştırıldığında, ALA ve trolox gruplarındaki oksidatif stres parametrelerindeki azalmalar istatistikî olarak önemli (P<0.05) bulundu. Bu çalışmanın sonuçlarına göre üreme mevsimi dışında Pırlak koç sperma sulandırıcısına ilave edilen ALA ve troloxun çözüm sonu spermatolojik parametreleri, oksidatif stresi ve DNA hasarına karşı koruyucu bir etki gösterdiği kanaatine varıldı.

Keywords

• Alfa lipoik asit,sezon dışı üreme,koç spermatozoon,trolox

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