Publication: Kronik psikolojik stres ile oluşturulmuş testis hasarı üzerine nöropeptit W’nin olası koruyucu etkilerinin morfolojik ve biyokimyasal olarak incelenmesi
Abstract
Amaç: Bu çalışmanın amacı, sıçanlarda kronik öngörülemeyen psikolojik stres (KS) modeli ile oluşturulmuş testis hasarı üzerinde Nöropeptit W’nin (NPW), olası koruyucu etkilerini değerlendirmektir. Bu kapsamda, testis morfolojisi, sperm parametreleri ve spermatogenezi düzenleyen hormonlar ve gen ekspresyonları, histolojik, immünohistokimyasal, biyokimyasal ve moleküler yöntemlerle incelendi. Gereç ve Yöntem: Sprague Dawley cinsi erkek sıçanlara, KS (avcı sesi, kedi kumu, yanıp sönen ışık, ıslak yatak, yiyecek yoksunluğu, eğik yatak ve ters ışık/ karanlık döngüsü) 40 gün boyunca uygulandı. Her stres uygulamasından önce de NPW (1 μg/ kg, ip.) verildi. Anksiyete düzeyleri kuyruk süspansiyon testiyle değerlendirildi. Kan serumunda, kortikosteron (CORT), adrenokortikotropik hormon (ACTH), testosteron, luteinize edici hormon (LH) ve folikül uyarıcı hormon (FSH) düzeyleri ölçüldü. Epididimal sperm parametreleri (konsantrasyon, canlılık, hareketlilik ve morfoloji) değerlendirildi. Histolojik olarak, testis hasarı, hücre proliferasyonu, apoptozis, oksidatif stres (NOX-2 pozitif hücreler) ve kan-testis bariyer proteini (Zonula okludens-ZO-1) değerlendirildi. Kan-testis bariyer proteinleri olan klaudin-11 ve okludin miktarları Western blot tekniği ile ölçüldü. Testis geçiş proteini-1 (TNP-1), klaudin-11 ve ZO-1 gen ifadeleri RT-qPCR yöntemiyle değerlendirildi. Testiste, malondialdehit (MDA), glutatyon (GSH) ve miyeloperoksidaz (MPO) düzeyleri biyokimyasal yöntemlerle ölçüldü. Tüm parametreler istatistiksel olarak analiz edildi. Bulgular: KS anksiyete seviyesini artırdı, kortizol ve ACTH düzeyini yükseltti, testosteron, LH ve FSH seviyelerini düşürdü ve sperm parametrelerini değiştirdi. Seminifer tübüllerde hasar, hücre proliferasyonunda, ZO-1 immünreaktivitesinde düşüş, apoptotik ve NOX-2 pozitif hücrelerde artış gözlendi. Klaudin-11 ve okludin proteinlerinin seviyesi ve TNP-1, klaudin-11 ve ZO-1 genlerinin ifadeleri azaldı. MDA ve MPO seviyelerinde artış olurken, GSH seviyesinde değişiklik gözlenmedi. NPW uygulaması tüm bu histolojik ve biyokimyasal bulguları düzeltti. Sonuç: KS, oksidatif testis hasarına, kan-testis bariyerinin ve sperm parametrelerinin bozulmasına neden olur. NPW, hormonal aktiviteyi ve kan-testis bariyerini düzenleyerek ve oksidatif stresi inhibe ederek testis hasarını önler ve sperm parametrelerini iyileştirir.
Objective: The aim of this study is to evaluate the possible protective effects of NPW on testicular damage induced by chronic unpredictable mild psychological stress (CUMS) model in rats. In this context, testicular morphology, sperm parameters, and hormones and gene expressions that regulate spermatogenesis were examined by histological, immunohistochemical, biochemical and molecular methods. Materials and Methods: Sprague Dawley male rats were exposed to CUMS (hunter sound, cat litter, flashing light, wet bedding, food deprivation, tilted bedding and reverse light/ dark cycle) for 40 days. NPW (1 μg/ kg, ip) was given before each stress application. Anxiety levels were assessed by tail suspension test. Corticosterone (CORT), adrenocorticotropic hormone (ACTH), testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels were measured in blood serum. Epididymal sperm parameters (concentration, viability, motility and morphology) were assessed. Histologically, testicular damage, cell proliferation, apoptozis, oxidative stress (NOX-2 positive cells) and blood-testis barrier (BTB) protein zonula occludens-1 (ZO-1) were evaluated. The BTB proteins claudin-11 and occludin were measured by Western blotting technique. Testis transition protein-1 (TNP-1), claudin-11 and zonula occludens (ZO)-1 gene expressions were evaluated by RT-qPCR. Testicular, malondialdehyde (MDA), glutathione (GSH) and myeloperoxidase (MPO) levels were measured by biochemical methods. Results: CUMS increased anxiety and cortisol and ACTH levels, decreased testosterone, LH and FSH levels and changed sperm parameters. Damage to seminiferous tubules, decrease in cellproliferation ZO-1 immunoreactivity, and increase in apoptotic and NOX-2 positive cells were observed. Claudin-11 and occludin protein’s levels, and the expressions of TNP-1, Claudin-11, and ZO-1 genes were decreased. Biochemical analysis showed an increase in MDA and MPO levels, but no changes in GSH level. NPW treatment ameliorated all these histological and biochemical findings. Conclusion: CUMS causes oxidative testicular damage, disruption of the blood-testis barrier and sperm parameters. NPW prevents testicular damage and improves sperm parameters by regulating hormonal activity and the blood-testis barrier and inhibiting oxidative stress.
Objective: The aim of this study is to evaluate the possible protective effects of NPW on testicular damage induced by chronic unpredictable mild psychological stress (CUMS) model in rats. In this context, testicular morphology, sperm parameters, and hormones and gene expressions that regulate spermatogenesis were examined by histological, immunohistochemical, biochemical and molecular methods. Materials and Methods: Sprague Dawley male rats were exposed to CUMS (hunter sound, cat litter, flashing light, wet bedding, food deprivation, tilted bedding and reverse light/ dark cycle) for 40 days. NPW (1 μg/ kg, ip) was given before each stress application. Anxiety levels were assessed by tail suspension test. Corticosterone (CORT), adrenocorticotropic hormone (ACTH), testosterone, luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels were measured in blood serum. Epididymal sperm parameters (concentration, viability, motility and morphology) were assessed. Histologically, testicular damage, cell proliferation, apoptozis, oxidative stress (NOX-2 positive cells) and blood-testis barrier (BTB) protein zonula occludens-1 (ZO-1) were evaluated. The BTB proteins claudin-11 and occludin were measured by Western blotting technique. Testis transition protein-1 (TNP-1), claudin-11 and zonula occludens (ZO)-1 gene expressions were evaluated by RT-qPCR. Testicular, malondialdehyde (MDA), glutathione (GSH) and myeloperoxidase (MPO) levels were measured by biochemical methods. Results: CUMS increased anxiety and cortisol and ACTH levels, decreased testosterone, LH and FSH levels and changed sperm parameters. Damage to seminiferous tubules, decrease in cellproliferation ZO-1 immunoreactivity, and increase in apoptotic and NOX-2 positive cells were observed. Claudin-11 and occludin protein’s levels, and the expressions of TNP-1, Claudin-11, and ZO-1 genes were decreased. Biochemical analysis showed an increase in MDA and MPO levels, but no changes in GSH level. NPW treatment ameliorated all these histological and biochemical findings. Conclusion: CUMS causes oxidative testicular damage, disruption of the blood-testis barrier and sperm parameters. NPW prevents testicular damage and improves sperm parameters by regulating hormonal activity and the blood-testis barrier and inhibiting oxidative stress.