Akademik Veri Yönetim Sistemi
Tezin Türü: Yüksek Lisans
Tezin Yürütüldüğü Kurum: Pamukkale Üniversitesi, Fen-Edebiyat Fakültesi, Biyoloji Bölümü, Türkiye
Tezin Onay Tarihi: 2012
Tezin Dili: Türkçe
Öğrenci: Gurbet Çelik
Danışman: Alaattin Şen
Özet:
The purpose of this project is to elucidate effects of ellagic acid on
xenobiotic metabolism and its drug-diet ınteraction potentials using
molecular approaches. For this purpose, ellagic acid was injected to
rats in two different doses and then their hepatic microsomal
7-ethoxyresorufin O-deethylase (CYP1A1),aminopyrene N- demethylase
(CYP2A, 2C), aniline 4-hydroxylase (CYP2E1), caffeine N-demethylase
(CYP1A2), dibenzofluoroscein demethylase (CYP19-aromatase), erythromycin
N-demethylase (CYP3A1), ethylmorphine N-demethylase (CYP2E1), caffeine
N-demethylase (CYP1A2), N-nitrosodimethylamine N-demethylase (CYP2E1),
methoxyresorufin O-demethylation (CYP1A2), benzyloxyresorufin
O-dealkylatase (CYP2B9), pentyloxyresorufin O-dealkylatase (CYP2B10),
benzphetamine N-demethylase (CYP2B), cytosolic glutathione S-transferase
(GST) and quinone reductase (NQO1) activities activities are going to
be determined.In order to determine the molecular mechanism of actions
of ellagic acid gene expression levels of xenobiotic metabolizing
enzymes given above was determined with the semi quantitative, RT-PCR.
Expression levels of xenobiotic metabolizing enzymes given above at the
protein level was be studied by Western blotting using specific
antibodies against individual isoforms. Cytotoxic effects of ellagic
acid on non cancer cell lines (293 and 3T3) and cancer cells lines
(breast cancer cell line MCF-7 and lung cell lines PC-14) was be
compared. Tumor suppressor genes such as PTEN and p53 and was be studied
in those cell lines with Western blotting to clarify the underlying
mechanism of cytotoxic actions observed with ellagic acid. In addition,
effect of ellagic acid on antioxidant enzymes (catalase and glutathione
peroxidase) and aspartate and alanine aminotransferases and was be
determined to study the oxidant, antioxidant possible toxic properties
of extracts and pure constituents. Although the CYP2B and CYP2E enzyme
activities were decreased, the activities of all other enzymes were not
changed significantly with 10 mg/kg EA injection. In addition,
Western-blot and qRT-PCR results clearly corroborated the above enzyme
expressions. On the other hand, while the NQO1, CAT, GPX, GSTs enzyme
activities were increased significantly, CYP450 1A1, 2B1, 2B4, 2B9,
2B10, 2C6, 2E1 and 19 enzyme activities were reduced significantly by an
i.p. administration of 30 mg/kg ellagic acid. In addition, Western-blot
and RT-PCR results markedly indicated that CYP2B, 2C6, 2E1 and 19
protein and mRNA levels were substantially decreased by 30 mg/kg dose of
EA, but the CYP1A protein and mRNA levels were not changed. CYP3A
enzyme activity, protein and mRNA levels were not altered by neither 10
nor 30 mg/kg ellagic acid. As a result, ellagic acid altered xeonobiotic
metabolism by cause change in the expression of CYP450 enzymes which
has an active role in the xenobiotic metabolism.