ENDOTOKSİN/D-GALAKTOZAMİN (LPS/GaIN) VERİLEN SIÇANLARIN ERİTROSİT ZARLARINDA LİPİT VE PROTEİN DEĞİŞİKLİKLERİ

Amaç: Endotoksin/D-galaktozamin (LPS/GaIN)’in sıçan eritrosit zarı üzerine etkilerini aydınlatmak. Gereç ve yöntem: Erkek Wistar sıçanlar, intraperitoneal 200 mg/kg D-galaktozamin (GaIN) ve 200 µg/ kg lipopolisakkarit (LPS) uygulamasından 18 saat sonra kesilerek ultrasantrifüj yöntemi ile eritrosit zarı (ghost) elde edildi. Sıçanların eritrosit zarlarında, malondialdehit (MDA), sülfidril (-SH) ve protein karbonil (PC) içeriği ve Na+ ,K+ -ATPaz aktivitesi spektrofotometrik olarak ölçüldü. MDA ve PC sıçanların plazmalarında da tayin edildi. Bunun yanısıra, eritrosit zar iskeletinin proteinlerindeki değişiklikler sodyum dodesil sülfat gel elektroforezi (SDS-PAGE) tekniği ile saptandı. Bulgular: LPS/GaIN uygulamasının, erkek Wistar sıçanlarda, serum alanin transaminaz (ALT) ve aspartat transaminaz (AST) ölçümleri sonucu, hepatit oluşturduğu belirlendi. Sıçanların plazma MDA düzeylerinde artış bulunurken, eritrosit zarlarında MDA ve PC düzeylerinin değişmediği gözlendi. Bununla birlikte sıçan eritrosit zarlarının –SH içeriği ve Na+ ,K+ -ATPaz aktivitesinde azalma olduğu saptandı. Eritrosit zar iskeleti proteinlerinin SDS-PAGE örneklerinde ise, proteinlerin bir kısmında, özellikle de, α-spektrin ve β-spektrin band yoğunluklarında azalmalar olduğu gözlendi. Sonuç: Bulgularımız, Na+ ,K+ -ATPaz aktivitesindeki azalmadan zar –SH içeriğindeki azalmanın sorumlu olabileceğini düşündürmektedir. Bunun yanısıra, eritrosit zar iskeletinin proteinlerinde görülen değişiklikler, hücre zar yapısında ve dolayısıyla fonksiyonlarında anormalliklere neden olarak LPS/GaIN hepatotoksisitesine katkıda bulunabilir.

Anahtar Kelimeler:

Lipit/protein oksidasyonu, zar iskelet proteinleri, LPS/GaIN, eritrosit zarı, sıçan

THE ALTERATIONS IN LIPID AND PROTEINS IN ERYTHROCYTE MEMBRANES OF ENDOTOXIN/DGALACTOSAMINE (LPS/GaIN)-ADMINISTERED RATS

Objective: To clarify the effects of endotoxin/D-galactosamine (LPS/GaIN) on erythrocyte membranes (ghost) of rats. Materials and methods: After 18 hours of intraperitoneal coadministration of 200 mg/kg D-galactosamine (GaIN) and 200 µg/kg lipopolysaccharide (LPS), male Wistar rats were decapitated and ghost was prepared by ultracentrifugation. Malondialdehyde (MDA), sulfhydryl (-SH) and protein carbonyl (PC) contents and Na+ ,K+ -ATPase activity were measured in ghost, spectrophotometrically. MDA and PC were measured in also plasma of rats. Furthermore, sodium dodesyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE) technique was used for the determination of the modifications in cytoskeletal ghost proteins . Results: LPS/ GaIN administration resulted in hepatitis in male Wistar rats as evaluated by assessments of serum alanine transaminase (ALT) and aspartate transaminase (AST) activities. While plasma MDA levels were determined to increase, ghost MDA and PC contents were observed unchanged. However, -SH content as well as Na+ ,K+ -ATPase activity of ghost were found decreased. As shown by the electrophoretic gel patterns, significant reduction in the intensities of some cytoskelatal membrane proteins, especially in α spectrin and β–spectrin, was observed in erythroctes of LPS/ GaIN-administered rats. Conclusion: From these findings, we concluded that reduced Na+ ,K+ -ATPase activity may be associated with the diminished – SH content in ghost of LPS/GaIN-administered rats. Furthermore, alterations seen in cytoskeletal proteins of ghost may participate to LPS/GaIN-induced hepatotoxicity by causing the deformation of cell membrane structure and abnormality of membrane functions, ultimately.

Keywords:

Lipid/protein oxidation, cytoskeletal proteins, LPS/GaIN, ghost, rat,

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