Koç Sperma Akrozom Bütünlüğünün Seminal Plazma Homosistein ve Nesfatin-1 Düzeyleri ile İlişkili Olarak İncelenmesi

Abstract

Bu çalışmada homosistein, nesfatin-1 düzeyleri ile akrozom bütünlüğü arasındaki ilişkinin araştırılması amaçlanmıştır. Ejakülatlar, suni vajen yöntemiyle altı yetişkin koçtan alındı ve toplanan ejakülatlar beş gruba bölünerek 37 °C'de sulandırıldı. Sperma sulandırılmasında kolesterol yüklü siklodekstrin (CLC) 1.5 ve 2.5 mg, kolesterol yüklü 7-dehidrokolesterol (7-DCLC) 1.5 ve 2.5 mg içeren veya katkı maddesi içermeyen (kontrol) sulandırıcılar kullanıldı. Tüm gruplardaki sperma örnekleri 5 °C' de soğutulduktan sonra sıvı nitrojen buharında (-110~-120° C) donduruldu. Sıvı nitrojen içinde saklanan sperma örnekleri incelenmek için 38 °C ve 30 saniyede çözdürüldü. Akrozom bütünlüğü, FITC-PNA boyama ile incelendi. Homosistein (HCY) ve nesfatin-1 testleri ELISA yöntemi ile yapıldı. CLC 1.5 ve 2.5 mg gruplarında homosistein seviyeleri kontrol grubuna (1.36±0.9) göre daha düşük bulundu (0.67±0.11, 0.61±0.26) (p <0.05). Nesfatin-1 düzeylerinde gruplar arasında istatistiksel olarak fark gözlenmedi (p> 0.05). 2.5 mg CLC'de akrozom bütünlüğü bozulmamış sperma hücrelerinin oranları (65.88±2.84) kontrol grubuna (52.38±2.97) göre daha yüksek bulundu (p<0.05). Sperma akrozom bütünlüğü, çözüm sonu koç spermasında HCY seviyesi (r =-,630) ile negatif, nesfatin-1 seviyeleri (r =, 460) ile pozitif korelasyon gösterdi. Sonuç olarak, CLC'nin akrozom bütünlüğü üzerinde kriyoprotektif bir etki gösterdiği ve HCY seviyesi üzerinde faydalı olduğu belirlenmiştir.

Keywords

• Akrozom bütünlüğü
• Siklodekstrin
• 7-dehidrokolesterol
• Homosistein
• Nesfatin-1

Investigation of Ram Sperm Acrosome Integrity in Relation with Seminal Plasma Homocysteine and Nesfatin-1 Levels

Abstract

This study aimed to investigate the relationship between homocysteine, nesfatin-1 levels with acrosome integrity. Ejaculates were collected from six adult rams via artificial vagina, divided into five groups and diluted at 37 °C. The extenders, which contained cholesterol loaded cyclodextrin (CLC) 1.5 and 2.5 mg, cholesterol loaded 7-dehydrocholesterol (7-DCLC) 1.5 and 2.5 mg or no additive (control), were used for sperm dilution. Semen samples of all groups were cooled at 5°C and then frozen in liquid nitrogen vapor (-110~-120°C). Semen samples stored in liquid nitrogen then were thawed at 38°C for 30 seconds prior to examination. Acrosome integrity was examined with FITC-PNA staining. Homocysteine (HCY) and nesfatin-1 assays were performed with ELISA method. In the CLC 1.5 and 2.5 mg groups homocysteine levels were lower (0.67±0.11;0.61±0.26) compared to control group (1.36±0.9) (p<0.05). No statistical differences were observed between groups in nesfatin-1 levels (p>0.05). In CLC 2.5 mg, the values of spermatozoa with intact acrosome membrane (65.88±2.84) were higher (p<0.05) than in control group (52.38±2.97). Sperm acrosome integrity was negatively correlated with HCY level (r =-,630) and positive correlated with nesfatin-1 levels (r =,460) in thawed ram sperm. In conclusion, CLC has a cryoprotective effect on acrosome integrity and found to have beneficial effects on HCY level.

Keywords

• Acrosome integrity
• Cyclodextrin
• 7-dehydrocholesterol
• Homocysteine
• Nesfatin-1

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