Pelophylax ridibundus'un yumurta jelinin fizikokimyasal karakterizasyonu ve melanom kanseri için biyolojik etkinliğinin belirlenmesi
Yükleniyor...
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Aksaray Ünversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu tez çalışmasında test organizma olarak seçilen, Ranidae familyasına ait bir amfibi türü olan Pelophylax ridibundus'un (P. ridibundus) embriyolarının beslenmesi, gelişmesi ve parazitlerden korunması için salgıladıkları jel yapısı ilk kez detaylı bir şekilde karakterize edilmiş ve literatüre kazandırılmıştır. Kozmopolit ve kolay kültüre edilebilir olması nedeniyle seçilen P. ridibundus'un yumurta jel yapısının fizikokimyasal analizleri (FT-IR, TGA, XRD ve elemental analiz), yapının tamamen karbohidrat ve protein yapıda olduğunu ortaya koymuştur. 96 µg/mL karbohidrat içerdiği tespit edilen yumurta jelinin, yapılan HPLC analizi ile profilinde Glikoz, Galaktoz, Mannoz ve Ksiloz varlığı tespit edilmiştir. LC-MS/MS analizi ile yapıda tespit edilen 2,4 µg/µL proteinin ise profili Uniprot/Swissprot veritabanı verilerine göre belirlenmiştir. Tespit edilen 12 proteinden sialik asit bağlı lektin proteini, tümör hücrelerine yönelik bir makrofaj sistemi yoluyla tümör büyümesini inhibe edebilme özelliği sayesinde dikkat çekmektedir. Kirli sucul ortam şartlarında amfibi embriyosunu koruyan, büyümesine ve gelişmesine olanak sağlayarak güçlü bir bariyer görevi gören yumurta jel yapısının, doğada bu zorlu ortam şartlarında özellikle UV ışınlarından embriyoyu koruması fonksiyonlarından esinlenerek, deri üzerine olumlu etkilerinin olabileceği ilk kez bu tez çalışmasında hipotez edilmiştir. Yüksek metastatik indeksi ile en agresif cilt kanseri olan melanom üzerine yaptığımız çalışmalarda amfibi yumurta jelinin bir biyomalzeme olarak güçlü biyolojik aktiviteye sahip olabileceği bu tez çalışması kapsamında ön görülmüştür. MTT analizi ve eş zamanlı ölçüm imkânı sunan xCELLigence cihazı ile yapılan hücre çalışmaları ile P. ridibundus yumurta jelinin 5 mg/mL selektif dozunun melanoma kanser hücrelerine (A375) antikanser bir etki gösterirken (canlılık < %30), sağlıklı insan deri hücrelerine sitotoksisite göstermediği (canlılık > %70) belirlenmiştir. Bu tez çalışmasında ilk kez doğadan gelen bir mucize olan amfibi yumurta jelinin melanom hücreleri üzerinde etkin bir biyoaktivite sergilediği ve potansiyel bir tedavi yöntemi olabileceği net olarak ortaya konmuştur.
In this thesis, the jelly structure, excreted for feeding, growing, and protecting the embryos of the selected test organism Pelophylax ridibundus (P. ridibundus), an amphibian species belonging to Ranidae family, was characterized in detail for the first time and gained to the literature. Physicochemical analysis (FT-IR, TGA, XRD, SEM-EDX, and elemental analysis) of the egg jelly structure of P. ridibundus, selected due to being cosmopolitan and ease of culture, demonstrated that the structure consists entirely of carbohydrate and protein. Through HPLC analysis, it was found that the egg jelly, identified to have 96 µg/mL carbohydrate, includes glucose, mannose, and xylose in its profile. 2,4 µg/µL protein was detected in the structure by LC-MS/MS analysis, and the protein profile was determined according to the Uniprot/Swissprote database data. Sialic acid-binding lectin, one of the identified 12 proteins, draws attention because of its ability to inhibit tumor growth via the way of the macrophage system against tumor cells. It was hypothesized for the first time in this study that the egg jelly, protecting the embryo of Amphibia in polluted aquatic conditions and acting the role of a strong barrier by contributing to its growth and improvement, may have positive effects on the skin, as inspired particularly by the protective functions of UV lights on the embryo in these challenging nature conditions. It was predicted within the framework of the current research study that the egg jelly of Amphibia could have a strong biological activity as a biomaterial discussed in the research on melanoma, the most aggressive skin cancer with a high metastatic index. Through cell studies carried out with MTT analysis and xCELLigence device enabling synchronous measurement, it was identified that while 5 mg/mL selective dose of egg jelly of P. ridibundus shows an anticancer effect (viability < %30) on melanoma cancer cells (A375), it does not show cytotoxicity (viability > %70) towards healthy human skin cells. It was clearly demonstrated for the first time in this thesis study that amphibian egg jelly, a natural miracle, exerts efficient bioactivity on melanoma cells and can be a potential treatment.
In this thesis, the jelly structure, excreted for feeding, growing, and protecting the embryos of the selected test organism Pelophylax ridibundus (P. ridibundus), an amphibian species belonging to Ranidae family, was characterized in detail for the first time and gained to the literature. Physicochemical analysis (FT-IR, TGA, XRD, SEM-EDX, and elemental analysis) of the egg jelly structure of P. ridibundus, selected due to being cosmopolitan and ease of culture, demonstrated that the structure consists entirely of carbohydrate and protein. Through HPLC analysis, it was found that the egg jelly, identified to have 96 µg/mL carbohydrate, includes glucose, mannose, and xylose in its profile. 2,4 µg/µL protein was detected in the structure by LC-MS/MS analysis, and the protein profile was determined according to the Uniprot/Swissprote database data. Sialic acid-binding lectin, one of the identified 12 proteins, draws attention because of its ability to inhibit tumor growth via the way of the macrophage system against tumor cells. It was hypothesized for the first time in this study that the egg jelly, protecting the embryo of Amphibia in polluted aquatic conditions and acting the role of a strong barrier by contributing to its growth and improvement, may have positive effects on the skin, as inspired particularly by the protective functions of UV lights on the embryo in these challenging nature conditions. It was predicted within the framework of the current research study that the egg jelly of Amphibia could have a strong biological activity as a biomaterial discussed in the research on melanoma, the most aggressive skin cancer with a high metastatic index. Through cell studies carried out with MTT analysis and xCELLigence device enabling synchronous measurement, it was identified that while 5 mg/mL selective dose of egg jelly of P. ridibundus shows an anticancer effect (viability < %30) on melanoma cancer cells (A375), it does not show cytotoxicity (viability > %70) towards healthy human skin cells. It was clearly demonstrated for the first time in this thesis study that amphibian egg jelly, a natural miracle, exerts efficient bioactivity on melanoma cells and can be a potential treatment.
Açıklama
Anahtar Kelimeler
Amfibi Yumurta Jeli, Karakterizasyon, Melanom Kanseri, Pelophylax ridibundus, Toksisite.
Kaynak
Aksaray Üniversitesi Fen Bilimleri Enstitüsü
