Metotreksatın neden olduğu testis hasarına karşı pirolidin ditiyokarbamatın koruyucu etkisinin incelenmesi
Küçük Resim Yok
Tarih
2019
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Trakya Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Metotreksat (MTX), klinikte yaygın kullanımı olan antikanser bir ilaçtır. En önemli yan etkilerinden biri, aktif çoğalan hücrelerin bulunduğu testislerde hasara neden olmasıdır. Pirolidin ditiyokarbamat (PDTC), güçlü bir antioksidan olup çeşitli ajanların neden olduğu testis hasarında histopatolojiyi düzelttiği bildirilmiştir. Bu çalışmada; MTX'in neden olduğu testis hasarı üzerine PDTC etkisinin, histolojik ve immünohistokimyasal olarak araştırılması planlanmıştır. Çalışmamızda, 40 adet Wistar albino erkek sıçan dört eşit gruba ayrıldı; Kontrol grubuna serum fizyolojik, PDTC ve MTX+PDTC grubuna, serum fizyolojikte çözülerek 100 mg/kg PDTC günlük toplam doz iki eşit doza bölünerek intraperitoneal (ip.) yol ile 10 gün boyunca verildi. MTX ve MTX+PDTC grubuna, 6. gün tek doz 20 mg/kg MTX ip. olarak verildi. Sıçanlar, PDTC'nin son dozundan 24 saat sonra sakrifiye edilerek testis dokuları histolojik ve immünohistokimyasal değerlendirme için alındı. İntrakardiak ponksiyon ile kan alınarak serum testosteron, FSH, LH, prokinetisin 2 düzeyleri saptandı. Hayvanların vücut ve testis ağırlıkları kaydedildi ve testis/vücut ağırlığı oranı hesaplandı. Sonrasında hematoksilen ve eozin boyalı preparatlardan seminifer tübül çapı, epitel kalınlığı ve intersitisyel saha genişliği ölçüldü. Histolojik hasar değerlendirmesi yapıldı ve immünohistokimyasal olarak NFkB, Nrf2, PK2 boyandı. Kontrol ve PDTC grupları arasında çalışılan parametreler arasında anlamlı bir fark bulunmazken, MTX verilen grupta; hayvanların vücut ve testis ağırlıkları, serum FSH, LH, testosteron düzeyleri, seminifer tübül çap ve germinal epitel kalınlığının kontrol ve PDTC gruplarına göre anlamlı olarak azaldığı, histolojik olarak hasarlı seminifer tübül sayısının, interstisyel saha genişliğinin ve PK2 seviyesinin ise, anlamlı olarak arttığı görüldü. Bundan başka, yine MTX grubunda; NFkB ve PK2 immünoreaktivitesinde kontrol ve PDTC gruplarına göre bir artış, Nrf2 immünoreaktivitesinde ise anlamlı azalma olduğu saptandı. Pirolidin ditiyokarbamat, MTX'in neden olduğu vücut ağırlığı, serum FSH, LH ve testosteron hormon düzeyi azalmasında anlamlı olarak düzelme sağlarken, histolojik olarak hasarlı seminifer tübül sayısı ve interstisyel saha genişlik artışında da anlamlı olarak azalma sağladı. Ek olarak PDTC, testis ağırlığında artış sağlarken, serum PK2 seviyelerini düşürdü. Ancak bu değişiklik istatistiksel olarak anlamlı bulunmadı. Ayrıca, artmış olan NFkB, PK2 immünoreaktivitelerinde azalmaya ve Nrf2 immünoreaktivitesinde ise artışa neden oldu. Sonuç olarak, PDTC tedavisinin MTX'in neden olduğu testis hasarını NFkB, Nrf2 ve PK2 sinyal yollarını etkileyerek azaltabileceği kanısındayız. Anahtar Kelimeler: Metotreksat, Pirolidin ditiyokarbamat, Testis, Nükleer faktör kappa B, Nükleer faktör eritroid 2 ilişkili faktör 2, Prokinetisin 2
Methotrexate (MTX) is an anticancer drug that is widely used in clinical practice. One of the most important side effects is the damage to the testes containing active proliferating cells. Pyrrolidine dithiocarbamate (PDTC) is a potent antioxidant and has been reported to improve histopathology in organ damage caused by various agents. In this study, we aimed to histologically and immunohistochemically investigate the protective effect of PDTC against MTX-induced testicular damage Forty Wistar albino male rats were divided into four equal groups: The control group; saline solution enjected intraperitoneally (ip.) for 10 days, PDTC and MTX+PDTC groups were injected PDTC which dissolved in saline solution, 100 mg/kg/day (daily total dose was divided into two equal doses) given ip. for 10 days. MTX and MTX+PDTC groups; single dose of 20 mg/kg MTX injected ip. on 6th day. Rats were sacrificed 24 hours after the last dose of PDTC and testis tissues were dissected out for histological and immunohistochemical evaluation. Serum testosterone, FSH, LH and prokineticin 2 (PK2) levels were determined by collecting blood samples via cardiac puncture. Change in body weight and testis weight of the animals were recorded and testis/body weight ratio was calculated. Afterwards, seminiferous tubule diameter, germinal epithelial thickness and interstitial area width were measured in hematoxylin-eosin stained preparations. Histological damage was assessed and NFkB, Nrf2, PK2 were stained immunohistochemically. Body and testicular weights, serum FSH, LH, testosterone, seminiferous tubule diameter, germinal epithelial thickness and PK2 levels were significantly decreased in the MTX group compared to control, and histologically damaged seminiferous tubule number and interstitial area width were significantly increased. Furthermore, there was an increase in NFkB and PK2 immunoreactivity in MTX group compared to control and PDTC groups, and a significant decrease in Nrf2 immunoreactivity. Pyrrolidine dithiocarbamate significantly increased MTX induced body weight, serum FSH, LH and testosterone hormone levels, while the number of histologically damaged seminiferous tubules and interstitial area width were reducing significantly. In addition, PDTC resulted in an increase in testicular weight while serum PK2 levels were reducing. However, this changes were not statistically significant. Furthermore, it caused a decrease in NFkB and PK2 immunoreactivity and an increase in Nrf2 immunoreactivity. In conclusion, we think that PDTC treatment may reduce MTX induced testicular damage by modulating NFkB, Nrf2 and PK2 pathways. Keywords: Methotrexate, Pyrrolidine dithiocarbamate, Testis, Nuclear factor kappa B, Nuclear factor erythroid 2 related factor 2, Prokineticin 2
Methotrexate (MTX) is an anticancer drug that is widely used in clinical practice. One of the most important side effects is the damage to the testes containing active proliferating cells. Pyrrolidine dithiocarbamate (PDTC) is a potent antioxidant and has been reported to improve histopathology in organ damage caused by various agents. In this study, we aimed to histologically and immunohistochemically investigate the protective effect of PDTC against MTX-induced testicular damage Forty Wistar albino male rats were divided into four equal groups: The control group; saline solution enjected intraperitoneally (ip.) for 10 days, PDTC and MTX+PDTC groups were injected PDTC which dissolved in saline solution, 100 mg/kg/day (daily total dose was divided into two equal doses) given ip. for 10 days. MTX and MTX+PDTC groups; single dose of 20 mg/kg MTX injected ip. on 6th day. Rats were sacrificed 24 hours after the last dose of PDTC and testis tissues were dissected out for histological and immunohistochemical evaluation. Serum testosterone, FSH, LH and prokineticin 2 (PK2) levels were determined by collecting blood samples via cardiac puncture. Change in body weight and testis weight of the animals were recorded and testis/body weight ratio was calculated. Afterwards, seminiferous tubule diameter, germinal epithelial thickness and interstitial area width were measured in hematoxylin-eosin stained preparations. Histological damage was assessed and NFkB, Nrf2, PK2 were stained immunohistochemically. Body and testicular weights, serum FSH, LH, testosterone, seminiferous tubule diameter, germinal epithelial thickness and PK2 levels were significantly decreased in the MTX group compared to control, and histologically damaged seminiferous tubule number and interstitial area width were significantly increased. Furthermore, there was an increase in NFkB and PK2 immunoreactivity in MTX group compared to control and PDTC groups, and a significant decrease in Nrf2 immunoreactivity. Pyrrolidine dithiocarbamate significantly increased MTX induced body weight, serum FSH, LH and testosterone hormone levels, while the number of histologically damaged seminiferous tubules and interstitial area width were reducing significantly. In addition, PDTC resulted in an increase in testicular weight while serum PK2 levels were reducing. However, this changes were not statistically significant. Furthermore, it caused a decrease in NFkB and PK2 immunoreactivity and an increase in Nrf2 immunoreactivity. In conclusion, we think that PDTC treatment may reduce MTX induced testicular damage by modulating NFkB, Nrf2 and PK2 pathways. Keywords: Methotrexate, Pyrrolidine dithiocarbamate, Testis, Nuclear factor kappa B, Nuclear factor erythroid 2 related factor 2, Prokineticin 2
Açıklama
Tıpta Uzmanlık
Anahtar Kelimeler
Histoloji ve Embriyoloji, Histology and Embryology
