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Maclura Pomifera Bitkisinden Izole Edilen Pomiferin Maddesinin Ratlarda Indometazin Ile Oluşturulan Gastrik Hasar Üzerine Etkilerinin Araştirilmasi

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Maclura Pomifera Bitkisinden Izole Edilen Pomiferin Maddesinin Ratlarda Indometazin Ile Oluşturulan Gastrik Hasar Üzerine Etkilerinin Araştirilmasi MACLURA POMİFERA BİTKİSİNDEN İZOLE EDİLEN POMİFERİN MADDESİNİN RATLARDA İNDOMETAZİN İLE OLUŞTURULAN GASTRİK HASAR ÜZERİNE ETKİLERİNİN ARAŞTIRILMASI İlyas BOZKURT Eczacılık Biyokimya Anabilim Dalı Tez Danışmanı Doç. Dr. Mesut Bünyami HALICI Yüksek Lisans Tezi – 2015 T.C ATATÜRK ÜNİVERSİTESİ SAĞLIK BİLİMLERİ ENSTİTÜSÜ MACLURA POMİFERA BİTKİSİNDEN İZOLE EDİLEN POMİFERİN MADDESİNİN RATLARDA İNDOMETAZİN İLE OLUŞTURULAN GASTRİK HASAR ÜZERİNE ETKİLERİNİN ARAŞTIRILMASI İlyas Bozkurt Eczacılık Biyokimya Anabilim Dalı Yüksek Lisans Tezi Tez Danışmanı Doç. Dr. Mesut Bünyami HALICI ERZURUM 2015 İÇİNDEKİLER TEŞEKKÜR ................................................................................................................... V ÖZET ............................................................................................................................ VI ABSTRACT ................................................................................................................. VII SİMGELER VE KISALTMALAR DİZİNİ ........................................................... VIII ŞEKİLLER DİZİNİ ....................................................................................................... X 1.GİRİŞ............................................................................................................................. 1 2.GENEL BİLGİLER ..................................................................................................... 3 2.1.Mide Anatomisi .......................................................................................................... 3 2.2.Midenin Yapısı ............................................................................................................ 3 2.3.Midenin Fonksiyonları ................................................................................................ 4 2.4.Midenin Fizyolojisi ..................................................................................................... 4 2.4.1.Mide Salgısı ............................................................................................................. 4 2.4.2.Mide Salgıları ve Fonksiyonları ............................................................................... 5 2.4.3.Mide Sekresyonunun Kontrolü ................................................................................ 6 2.5.Ülser ............................................................................................................................ 7 2.5.1.Epidemiyolojisi ........................................................................................................ 7 2.5.2.Etiyoloji .................................................................................................................... 8 2.5.3.Non-Steroid Antiinflamatuar İlaçlar ve Hasar Oluşum Mekanizması ................... 10 2.6.Midenin Ülser Savunması ve Mekanizması ............................................................. 11 2.7.Antioksidan Savunma Sistemi .................................................................................. 13 2.7.1.Antioksidanlar ........................................................................................................ 13 2.7.2.Serbest Radikal Kaynakları:................................................................................... 18 2.7.3.Serbest Radikallerin Etkileri .................................................................................. 22 2.7.4.Antioksidan Savunma Sistemleri ........................................................................... 26 2.8.Ülser ve Oksidatif Stres İlişkisi ................................................................................ 36 2.9.Maclura pomifera ..................................................................................................... 37 2.9.1.Maclura pomifera’nın Kimyasal Bileşeni ............................................................. 39 2.10.Pomiferin ................................................................................................................. 39 3.MATERYAL VE METOT ........................................................................................ 41 3.1.Deneylerde Kullanılan Kimyasallar .......................................................................... 41 3.2.Deneylerde Kullanılan Cihazlar ................................................................................ 41 3.3.Deney Materyalinin Temini ve Etken Maddenin İzolasyonu ................................... 41 3.4.Deney Hayvanlarının Temini .................................................................................... 43 III 3.5.Ratlarda İndometazin Uygulaması ile Deneysel Ülser Oluşturulması ..................... 43 3.5.1.Ülser Modeli .......................................................................................................... 43 3.6.Mide Dokusunun Makroskopik İncelenmesi ............................................................ 45 3.7.Mide Dokusunun Biyokimyasal İncelenmesi ........................................................... 45 3.7.1.Doku Homojenatlarının Hazırlanması: .................................................................. 46 3.7.2.Deneylerde Kullanılan Çözeltiler ve Hazırlanışları ............................................... 46 3.7.3.Lipid Peroksidasyon Seviyesinin Ölçümü: ............................................................ 49 3.7.4.Total Glutatyon (GSH) Miktarı Ölçümü: .............................................................. 50 3.7.5.Süperoksit Dismutaz (SOD) Aktivitesinin Ölçümü: .............................................. 51 3.7.6.Katalaz (KAT) Aktivitesinin Ölçümü: ................................................................... 52 3.8.İstatistiksel Analizler ................................................................................................ 53 4.BULGULAR ............................................................................................................... 54 4.1.Makroskobik Analiz .................................................................................................. 54 4.2.Biyokimyasal Analiz ................................................................................................. 56 4.2.1.LPO Seviyesi ......................................................................................................... 56 4.2.2.SOD Aktivitesi ....................................................................................................... 57 4.2.3.GSH Seviyesi ......................................................................................................... 58 4.2.4.KAT Aktivitesi ....................................................................................................... 60 5.TARTIŞMA ................................................................................................................ 62 6.SONUÇ VE ÖNERİLER ........................................................................................... 74 KAYNAKLAR .............................................................................................................. 75 EKLER ......................................................................................................................... 101 EK-1. ÖZGEÇMİŞ ..................................................................................................... 101 EK-2. ETİK KURUL ONAY FORMU ..................................................................... 102 IV TEŞEKKÜR Yüksek Lisans tezi olarak sunduğum bu çalışmada, lisansüstü eğitimim ve tez çalışmam boyunca yanımda olan, sınırsız desteğini sunan değerli danışmanım Sayın Doç. Dr. Mesut Bünyami HALICI hocama en derin saygılarımı ve şükranlarımı sunarım. Lisansüstü eğitimim süresince yanımda olan ve öğrendiklerim konusunda büyük emeğe sahip, hem tez çalışmamda hem de bu çalışmayı başarıyla sonuca ulaştırmamda yardımlarını esirgemeyen Veteriner Biyokimya A.B.D’ nın değerli hocalarından olan Sayın Yrd. Doç. Dr. Hüseyin Serkan EROL’ a, çalışmada kullandığım pomiferin maddesini saflaştırma ve tanımlamada büyük emeği olan Sayın Prof. Dr. Ahmet ÇAKIR’ a (Kilis 7 Aralık Üniversitesi), tez kapsamında deney hayvanlarının temini ve deneyin gerçekleştirilmesinde büyük emekleri olan Sayın ATADEM yönetimi ve değerli çalışanlarına, çalışmanın gerçekleşebilmesi için maddi desteklerini esirgemeyen, bu çalışmayı 213S114 nolu projeyle destekleyen TÜBİTAK’ a, ayrıca Sağlık Bilimleri Enstitüsü Müdürlüğü ve çalışanlarına ve hayatım boyunca gerek aldığım kararlar gerekse eğitimim boyunca her zaman beni destekleyen aileme teşekkürlerimi sunarım. İlyas BOZKURT V ÖZET Maclura pomifera Bitkisinden İzole Edilen Pomiferin Maddesinin Ratlarda İndometazin ile Oluşturulan Gastrik Hasar Üzerine Etkilerinin Araştırılması Amaç: Daha önceki çalışmalarda Maclura pomifera bitkisinin su ekstresinin gastrik hasar üzerine etkili olduğu bildirilmiş, bu çalışmada ise bu etkinliğe sahip olduğu düşünülen pomiferin maddesi saflaştırılarak, ratlarda indometazin ile oluşturulan gastrik hasar üzerine antiülser etkileri incelenmiştir. Materyal ve Metot: Çalışma için gerekli olan pomiferin Maclura pomifera bitkisinden İTK yöntemi ile saflaştırılarak, NMR yöntemi ile yapısı aydınlatıldı. Çalışma için, toplam 36 adet Sprague Dawley cinsi erkek rat temin edilerek, 6’şarlı 6 gruba ayrıldı. REF grubuna 25 mg/kg ranitidin (Ulcuran® 25 mg/ml flakon), PMF-100 grubuna 100 mg/kg, PMF-200 grubuna 200 mg/kg, PMF-300 grubuna 300 mg/kg pomiferin ve sağlıklı grup hariç tüm gruplara 25 mg/kg indometazin uygulandı. Ratlar, uygulamadan 6 saat sonra sakrifiye edilerek, midelerinde milimetrik kağıt
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