Characterization of aging of drug metabolizing enzymes and assessment of aging in gilthead seabream (sparus aurata) liver microsomes

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Tezin Türü: Yüksek Lisans

Tezin Yürütüldüğü Kurum: Pamukkale Üniversitesi, Fen-Edebiyat Fakültesi, Kimya Bölümü, Türkiye

Tezin Onay Tarihi: 2006

Tezin Dili: Türkçe

Öğrenci: Hatice Ardağ

Danışman: Alaattin Şen

Özet:

Cytochrome P450 (CYP) comprises a superfamily of hemoproteins that play a pivotal role in the activationand detoxification of xenobiotics such as drugs, pesticides, antioxidants, organic solvents, anesthetic agens, dyes,enviromental pollutants and food additives and endogenous compounds including steroids, bile acids, vitaminD3, fatty acids, prostaglandins, biogenic amines and retinoids. The cytochrome P450 (CYP450) family is themajor phase I metabolic enzyme system in the liver.In this study, gilthead seabream (Sparus aurata) fish liver microsomes and cytosol of different ages wereused as sample. The fish used in this study, Gilthead Seabream (Sparus Aurata), were bought from Pinar Fish inİzmir, Aegean coast of Turkey. In this study Gilthead Seabream (Sparus Aurata) Fish Liver Microsomes andcytosoles of different ages (ranging from 1.5 to 24 months) were used. Liver microsomes and cytosoles wereprepared according to the method of Arinc and Sen (1998). Liver microsomal Ethoxyresorufin O-deethylase(EROD), methoxyresorufin O-demethylase (MROD), penthyloxyresorufin O-depenthylase (PROD),benzyloxyresorufin O-debenzylase (BROD), aniline 4-hydroxylase (A4H), N-nitrosodimethylamine N-demethylase (NDMA-DE), aminopyrene N-demethylase (APND) ve erytromycin N-demethylase (ERND) weredetermined from this Gilthead Seabream (Sparus aurata) fish liver microsomes.A4H activity increased with aging. While 1.5 months of the fish activity is 0,002 ± 0,001 nmol/dakika/mgprotein, 24 months of the fish is 0,058 ± 0,002 nmol/min/mg protein. ERND activity increased in giltheadseabream (Sparus aurata) liver microsomes of different ages (ranging from 1.5 to 24 months). 1.5 months of thefish activity is 0,09 ± 0,006 nmol/dakika/mg protein, 24 months of the fish is 0,189 ± 0,008 nmol/min/mg protein.NDMA-ND activity increased in gilthead seabream (Sparus aurata) liver microsomes of different ages (rangingfrom 1.5 to 24 months). 1.5 months of the fish activity is 0,019 ± 0,007 nmol/min/mg protein, 24 months of thefish is 0,323 ± 0,014 nmol/min/mg protein.CN3D activity decreased in gilthead seabream (Sparus aurata) liver microsomes of different ages (rangingfrom 1.5 to 24 months). 1.5 months of the fish activity is 0,438 ± 0,014 nmol/min/mg protein, 24 months of thefish is 0,069 ± 0,012 nmol/min/mg protein. APND activity decreased in gilthead seabream (Sparus aurata) livermicrosomes of different ages (ranging from 1.5 to 24 months). 1.5 months of the fish activity is 0,944 ± 0,024nmol/min/mg protein, 24 months of the fish is 0,056 ± 0,011 nmol/min/mg protein.EROD activity increased in gilthead seabream (Sparus aurata) liver microsomes of different ages (rangingfrom 1.5 to 24 months). 9 months of the fish activity is 88,6 ± 1,11 pmol/min/mg protein, 24 months of the fish is998,4 ± 13,7 pmol/min/mg protein. 1.5 to 9 months of the fish liver activity were not detected. PROD activityincreased in gilthead seabream (Sparus aurata) liver microsomes of different ages (ranging from 1.5 to 24months). 15 months of the fish activity is 285,6 ± 20,9 pmol/dakika/mg protein, 24 months of the fish is 613,7 ±16,4 pmol/dakika/mg protein. 1.5 to 15 months of the fish liver activity were not detectable. MROD and BRODactivities were not detectable.Glutathione S-transferase (GST) were determined from this Gilthead Seabream (Sparus aurata) fish liverscytosoles. GST-CDNB activity increased in gilthead seabream (Sparus aurata) liver microsomes of differentages (ranging from 1.5 to 20 months). 1.5 months of the fish activity is 17,7 ± 0,5 pmol/dakika/mg protein, 20months of the fish is 690,9 ± 32,0 pmol/dakika/mg protein.