Özlem Özden AKKAYA, Artay YAĞCI, Murat TOSUN, Korhan ALTUNBAŞ

Fötal ve Neonatal Rat Beyin Dokusunda Bisphenol A’nın Notch1 İmmunohistokimyasal Ekspresyonu Üzerine Etkisi

Bisphenol A (BPA), tekrar kullanılabilir plastik kaplarda, gıda ve içecek kutularında, biberonlarda ve diş dolgularında kullanılan bir plastikleştiricidir. BPA’nın bu ürünlerden hidrolize olarak açığa çıkması ve bulunduğu kabın içerisindeki maddelere geçebilmesi canlılarda önemli sağlık problemlerine yol açar. En önemlisi BPA’nın plasental ve laktasyonal yolla geçerek henüz intrauterin dönemdeki veya laktasyonal evredeki canlının gelişimini etkileyebilmesidir. Notch sinyal yolağı hücrelerin farklılaşmasını, çoğalmasını ve canlı kalmasını düzenleyerek hücre kaderini belirler. Notch sinyal yolağı üyeleri pek çok organ gibi beyinde de ekspre olmaktadır. Notch sinyal yolağı üyelerinden Notch1 ventrikulus çevresinde [ventrikulus duvarı ve subventrikular zon (ventrikulusun altındaki bölge)], pleksus koroyideyus, subtansiya grizea ve substansiya alba, hipokampüs ve serebral damarlarda ekspre olur. Çalışmamızda fötal ve neonatal dönemlerde 5 farklı zaman aralığında BPA’nın Notch1 ekspresyonu üzerine etkisini araştırdık. Çalışmada Wistar ırkı ratlar kullanıldı (n=60). 5 farklı deney ve kontrol grubu oluşturuldu. Deney gruplarına 50 mg/kg/gün BPA, kontrol gruplarına ise taşıt madde [Susam yağı+etanol (SE), 9:1] uygulandı. Deneyin birinci kısmında Embriyonik (E)18-21, Postnatal (P) 0-3 ve P4-7 dönemlerinde uygulama yapıldı. E18-21 grubuna gebeliğin 18. gününden itibaren gebe annelere günlük olarak, intraperitoneal (i.p.), P0-3 ve P4-7 gruplarında ise yavrulara belirtilen uygulama aralıklarında subkutan (s.c.) BPA veya taşıt madde verildi. Tüm yavrular P7. günde sakrifiye edilerek beyin dokuları alındı. Deneyin ikinci kısmında E18-21, P0-3 dönemlerde birinci deneydekine benzer uygulama yapıldı. Uygulamanın bitmesini takiben E21. günde gebe ratlar sakrifiye edilerek fötusların beyin dokuları ve P3. günde yavru ratlar sakrifiye edilerek yavruların beyin dokuları alındı. Notch1 ekspresyonu immunohistokimya yöntemi ile değerlendirildi. Beyin dokusunda Notch1 pia mater, substansiya grizea, ventrikuluslar çevresi [ventrikulus duvarı ve subventrikular zon (ventrikulusun altındaki bölge)], pleksus koroyideyus ve kapillar endotellerinde ekspre oldu. Fötal ve neonatal dönemdeki BPA uygulamalarını takiben E21., P3. ve P7. günlerde farklı düzeylerde Notch1 ekspresyonu gözlendi. Sonuç olarak BPA’nın beyinde Notch1 ekspresyonları üzerine etkilerinin maruz kalınan döneme ve gelişim sürecine bağlı olarak değişebileceği görüldü.

Effects of Bispherol A on Notch1 Immunohistochemical Expression in Fetal and Neonatal Rat Brain

Bisphenol A (BPA) is a plasticizer found in reusable plastic containers, food and beverage cans, baby bottles and dental sealants. BPA exposure has become an important health concern based on its ability to “leach” from these products and penetrate the materials contained within them. Importantly, BPA can be transferred via placenta or lactation and this may affect the living body in intrauterine or lactation period. Notch signaling pathway to regulate cell fate by modulating differentiation, proliferation, and survival of cells. It has been reported that the members of Notch signaling are expressed in brain. Notch1 is a member of Notch signalling pathway, that expresses in subventricular zone, choroid plexus, grey matter, white matter, hippocampus and cerebral vessels of brain. In our study, we examined the effect of BPA on Notch1 expression in 5 different time intervals of fetal and neonatal periods in vivo. Wistar rats were used in this study (n=60). Five different experimental and control groups were formed. The experimental groups were treated with BPA at 50 mg/kg/day when control groups were treated with sesame oil and ethanol at 9:1 (vehicle). During the first part of the experiment, BPA or vehicle was applied to three groups at E 18-21, P 0-3 and P 4-7 periods. When BPA or vehicle were injected intraperitoneally to pregnant dams in E 18-21 group neonatal pups in P0-3 and P4-7 groups were given subcutaneous injections. The pups were sacrificed at the end of 7th day and their brain tissues were collected. During the second part of the experiment, similar applications with first experiment was performed. Following the application, pregnant dams were sacrificed and brain tissue of their fetuses were collected at E21st day and neonatal pups were sacrificed at P3rd day and their brains were collected. Notch1 expression was assessed by using immunohistochemistry. Notch1 was expressed in the pia mater, the grey matter, around ventricles (ventricular walls and sub ventricular zone) and in choroid and vascular plexus of brain. After BPA applications in fetal and neonatal periods, Notch1 expression was seen in different levels at E21st, P3rd and P7th days. In conclusion, the effects of BPA on Notch1 immunohistochemical expression in brain tissue, varies depends on exposure time and the developmental period during the exposure.

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