Varikosel olgularında spermin DNA fragmantasyonu ve immunohistokimyasal incelenmesi
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Tarih
2024
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Yayıncı
Dicle Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Özet Amaç: Bu çalışmanın temel amacı, varikosel olgularında spermin DNA fragmentasyon düzeylerini değerlendirmek ve immunhistokimyasal yöntemler kullanarak sperm hücrelerindeki potansiyel değişiklikleri saptamaktır. Araştırma, varikoselin spermatogenezi ve sperm DNA'sı üzerindeki etkilerini mikroskopik düzeyde incelemeyi hedeflemektedir. Gereç ve Yöntem: Bu çalışmada, 25-40 yaş aralığındaki infertil erkeklerin semen örnekleri, varikosel varlığı ve diğer belirlenen faktörler açısından incelenmiş ve normozoospermik kontrol grubu ve varikosel teşhisi almış grup olarak iki gruba ayrılmıştır. Yüksek motiliteye sahip spermatozoaları ayırmak için direkt yüzdürme teknikleri uygulanmış, bu sayede DNA fragmentasyonu ve sperm morfolojisi detaylıca analiz edilmiştir. Sperm vitalitesi Eosin Y boyası ile, DNA fragmentasyonu Akridin Oranj boyama metoduyla ve sperm morfolojisi Spermac boyama kitleri kullanılarak değerlendirilmiştir. İmmünhistokimyasal boyama yöntemleriyle protein ekspresyonu görselleştirilmiş ve sonuçlar mikroskopi ile fotoğraflanarak kaydedilmiştir. Bulgular: Dicle Üniversitesi Tıp Fakültesi Üroloji Anabilim Dalı'nda yürütülen bu çalışmayla, varikosel tanısı konmuş ve sağlıklı kontrol grubu oluşturan erkeklerden alınan sperm örnekleri üzerinde gerçekleştirilmiştir. Bulgular, dinein ve aquaporin proteinlerinin, sperm hücrelerinde varikosel varlığının düşük ya da negatif ekspresyonu ile ilişkili olduğunu göstermektedir. İmmünohistokimyasal analizler, varikosel hastalarında dinein proteinlerinin ve aquaporin 3 ve 5'in sağlıklı bireylerle karşılaştırıldığında azalmış ya da eksik ifadesini ortaya koymuştur. Bu, sperm motilitesi ve su transfer mekanizmalarında olası bir bozulmaya işaret etmekte ve varikoselin erkek fertilitesi üzerindeki etkilerinin daha derinlemesine anlaşılmasına katkı sağlamaktadır. Sonuç: Bu çalışma, varikosel hastalığının spermatozoanın yapısal ve fonksiyonel integritesi üzerindeki olumsuz etkilerini açıkça ortaya koymuştur. Dinein ve aquaporin protein aileleri, spermatozoanın hareket kabiliyeti ve su dengesi için kritik öneme sahip olduğundan, bu proteinlerin düşük ekspresyonu ya da yokluğu, erkek infertilitesinin moleküler mekanizmalarına ışık tutmaktadır.
Abstract Aim: The primary objective of this study is to evaluate the levels of DNA fragmentation in sperm in cases of varicocele and to detect potential alterations in sperm cells using immunohistochemical methods. The research aims to microscopically examine the effects of varicocele on spermatogenesis and the integrity of sperm DNA. Materials and Methods: In this study, semen samples collected with consent from infertile males aged 25-40 were analyzed for the presence of varicocele and other identified factors and were divided into two groups: a normozoospermic control group and a group diagnosed with varicocele. Direct swim-up techniques were employed to separate highly motile spermatozoa, thereby facilitating a detailed analysis of DNA fragmentation and sperm morphology. Sperm viability was assessed using Eosin Y staining, DNA fragmentation was evaluated with Acridine Orange staining methods, and sperm morphology was examined using Spermac staining kits. Protein expression was visualized through immunohistochemical staining methods, and the results were recorded using high magnification photography. Results: This research was conducted on sperm samples collected from males diagnosed with varicocele and a healthy control group at the Urology Department of Dicle University Medical Faculty. The findings indicate that the presence of varicocele in sperm cells is associated with reduced or negative expression of dynein and aquaporin proteins. Immunohistochemical analyses have revealed a diminished or absent expression of dynein proteins and aquaporins 3 and 5 in varicocele patients compared to healthy individuals. This suggests a potential disruption in sperm motility and water transfer mechanisms, contributing to a deeper understanding of the effects of varicocele on male fertility. Conclusion: The outcomes of this study unequivocally reveal the deleterious effects of varicocele disease on the structural and functional integrity of spermatozoa. Given that the dynein and aquaporin protein families are crucial for the movement and fluid balance of spermatozoa, the reduced expression or absence of these proteins sheds light on the molecular mechanisms of male infertility.
Abstract Aim: The primary objective of this study is to evaluate the levels of DNA fragmentation in sperm in cases of varicocele and to detect potential alterations in sperm cells using immunohistochemical methods. The research aims to microscopically examine the effects of varicocele on spermatogenesis and the integrity of sperm DNA. Materials and Methods: In this study, semen samples collected with consent from infertile males aged 25-40 were analyzed for the presence of varicocele and other identified factors and were divided into two groups: a normozoospermic control group and a group diagnosed with varicocele. Direct swim-up techniques were employed to separate highly motile spermatozoa, thereby facilitating a detailed analysis of DNA fragmentation and sperm morphology. Sperm viability was assessed using Eosin Y staining, DNA fragmentation was evaluated with Acridine Orange staining methods, and sperm morphology was examined using Spermac staining kits. Protein expression was visualized through immunohistochemical staining methods, and the results were recorded using high magnification photography. Results: This research was conducted on sperm samples collected from males diagnosed with varicocele and a healthy control group at the Urology Department of Dicle University Medical Faculty. The findings indicate that the presence of varicocele in sperm cells is associated with reduced or negative expression of dynein and aquaporin proteins. Immunohistochemical analyses have revealed a diminished or absent expression of dynein proteins and aquaporins 3 and 5 in varicocele patients compared to healthy individuals. This suggests a potential disruption in sperm motility and water transfer mechanisms, contributing to a deeper understanding of the effects of varicocele on male fertility. Conclusion: The outcomes of this study unequivocally reveal the deleterious effects of varicocele disease on the structural and functional integrity of spermatozoa. Given that the dynein and aquaporin protein families are crucial for the movement and fluid balance of spermatozoa, the reduced expression or absence of these proteins sheds light on the molecular mechanisms of male infertility.
Açıklama
Anahtar Kelimeler
Sperm DNA fragmantasyonu, Immünhistokimya, Varikosel, Protein ekspresyonu, Erkek infertilitesi, Sperm DNA fragmentation, Immunohistochemistry, Varicocele, Protein expression, Male infertility
Kaynak
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Künye
Duran, S. Ç. (2024). Varikosel olgularında spermin DNA fragmantasyonu ve immunohistokimyasal incelenmesi. Yayımlanmamış doktora tezi, Dicle Üniversitesi, Diyarbakır.
