The effects of CAPE (caffeic acid phenethyl ester) addition on spermatological parameters, oxidative stress and DNA damage after frozen-thawed process in ram semen.

Abstract

The aim of this study is to investigate the effects of different doses caffeic acid phenethyl ester (CAPE) adding to extender on some spermatological parameters, oxidative stress and DNA damage during post-thawed of ram semen. Ejaculates were collected from five Merino rams using an artificial vagina ones a week and this process was repeated six times. Ejaculates were split into four aliquots and diluted to a final concentration of 150x106 spermatozoa/ml with the base extender containing antioxidant (10 µg/ml, 50 µg/ml and 100 µg/ml)and no additive (control). All samples were cooled to 4°C and equilibrated for 3 h then were loaded into 0.25 ml straws and frozen using a liqued nitrogen vapour and plunged into liquid nitrogen. In terms of subjective motility, the groups containing 50 and 100 μg / ml were found to be significantly superior (P <0.05) compared to the control group. In addition, in terms of mid-piece anomalies and total abnormal spermatozoon rate, the decrease in the group containing 100 μg / ml compared to the control group and in the groups containing 50 and 100 μg / ml in terms of tail anomalies was found to be significant (P <0.05). The increase in all antioxidant groups in terms of H + / E- ratio compared to the control group, decreases in the group containing 100 μg / ml CAPE compared to the control group in terms of tail length, tail DNA and tail moment, and the spermatozoon TAS levels in the groups containing 10 and 100 μg / ml CAPE compared to the control group. the increase was statistically significant (P <0.05). In conclusion, in our study, it was observed that the 100 μg / ml dose of CAPE in different doses added to the sperm in the storage of ram semen provided the best protection compared to the control group and other groups in terms of spermatozoon motility, abnormal spermatozoon rate, HOST-Eosin test rate, oxidative stress and DNA damage.

Keywords

• CAPE (caffeic acid phenethyl ester)
• frozen
• DNA damage
• ram
• oxidative stres

Koçlarda spermaya katılan CAPE'in (caffeic acid phenethyl ester) dondurma ve çözdürme sonrası spermatolojik parametreler, oksidatif stres ve DNA hasarı üzerine etkileri

Öz

Bu çalışmanın amacı koç sperma sulandırıcısına ilave edilen farklı dozlardaki caffeic acid phenethyl ester (CAPE) çözüm sonu spermatolojik parametreler, oksidatif stres ve DNA hasarı üzerine etkileri araştırıldı. Ejakulatlar beş baş Merinos koçtan haftada bir kez suni vajen yardımıyla toplandı ve bu işlem altı kere tekrarlandı. Ejakulatlar ml’de 150x106 spermatozoon olacak şekilde antioksidan içermeyen (kontrol) ve antioksidan içeren (10 µg/ml, 50 µg/ml ve 100 µg/ml) sulandırıcılar ile dört bölüme ayrıldı. Sulandırılan örnekler 0,25 ml’lik payetlerde 5 0C’de 3 saat ekilibrasyona tabi tutulduktan sonra sıvı azot buharında donduruldu. Subjektif motilite yönünden 50 ve 100 μg/ml içeren grupların, kontrol grubuna göre belirgin bir üstünlük sağladığı (P<0.05) gözlendi. Ayrıca orta kısım anomalileri ile toplam anormal spermatozoon oranı bakımından kontrol grubuna göre 100 μg/ml içeren gruptaki ve kuyruk kısmındaki anomaliler açısından 50 ve 100 μg/ml içeren gruplardaki azalma önemli (P<0.05) bulunmuştur. H+/E- oranı açısından kontrol grubuna göre tüm antioksidan gruplarındaki artış, tail lenght, tail DNA ve tail moment bakımından kontrol grubuna göre 100 μg/ml CAPE içeren gruptaki azalmalar ve kontrol grubuna göre spermatozoon TAS düzeylerinde 10 ve 100 μg/ml CAPE içeren gruplardaki artış istatistikî olarak önemli (P<0.05) bulundu. Sonuç olarak yaptığımız çalışmada koç spermanın saklanmasında spermaya katılan farklı dozlardaki CAPE'in 100 μg/ml dozu spermatozoon motilite, anormal spermatozoon oranı, HOST-Eozin test oranı, oksidatif stres ve DNA hasarı yönünden kontrol grubu ve diğer gruplara göre en iyi korumayı sağladığı görüldü.

Anahtar Kelimeler

• CAPE (caffeic acid phenethyl ester)
• dondurma
• DNA hasarı
• koç
• oksidatif stres

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