Endokrin bozucuların steroidojenez üzerine etkilerinin hormon analizleri ile belirlenmesi
Küçük Resim Yok
Dosyalar
Tarih
2021
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Endokrin bozucular (EB), sağlıklı organizmada veya gelecek nesillerinde advers sağlık etkilerine, endokrin fonksiyonlarda değişikliklere sebep olan ekzojen maddeler olarak tanımlanmaktadır. EB hormon reseptörleri ile etkileşerek agonist ya da antagonist etki gösterebileceği gibi steroid hormonların biyosentez ve metabolizma yolaklarında yer alan enzimlerle etkileşerek fizyolojik hormon düzeylerini değiştirmek yoluyla da etkilerini gösterebilmektedir. Bu tez çalışmasında endokrin bozucuların cinsiyet hormonlarının sentez yolağı olan steroidojenez üzerine etkilerinin araştırılması amacıyla OECD TG-456 test kılavuzunda belirtilen valide in vitro bir yöntemin laboratuvarımızda uygulanması ve deneyin sonucunda östradiol (E2) ve testosteron (T) düzeylerinin belirlenmesi amacıyla ELİSA ve LC MS/MS yöntemlerinin kullanılması, her iki analiz yöntemi ile elde edilen verilerin karşılaştırılması amaçlanmıştır. Yöntemde pozitif kontrol olarak (enzim indükleyici etki) OECD kılavuzunda belirtildiği gibi forskolin, negatif kontrol olarak (enzim inhibitörü etki) prokloraz, steroidojenez döngüsünde E2 ve T üzerine etkisi bilinen referans maddelerden ketakonozol, letrozol ve genistein kullanılarak yöntem(ler)in kısmi validasyonları yapılmış ve ölçüm performansları karşılaştırılmıştır. Her iki yöntem arasında fark olup olmadığı istatistiksel olarak değerlendirilmiştir. Takiben hormon bağımlı meme kanseri tedavisinde aromataz enzim inhibitörü olarak kullanılması önerilecek, yeni sentezlenmiş ilaç etkin madde adayları olan 2-Metil-1-H-indol-3-karboksialdehit (4-klorofenil) hidrazon (M6) ve 2-metilindol-3-karboksialdehit fenil hidrazon (M20) ile bunların model bileşiği olan melatoninin tüm steroidojenez yolağı üzerindeki etkileri de H295R steroidojenez yöntemi uygulanması sonrasında hormon analizleri her iki yöntemle yapılarak araştırılmıştır. Sonuç olarak uygulanan iki yöntem avantaj ve dezavantajları açısından değerlendirilmiştir.
Endocrine disruptors (EB) are defined as exogenous substances that cause adverse health effects and changes in endocrine functions in healthy organisms or in their future generations. EB can act as an agonist or antagonist by interacting with hormone receptors, or it can also exert its effects by changing physiological hormone levels by interacting with enzymes involved in the biosynthesis and metabolism pathways of steroid hormones. In this thesis study, in order to investigate the effects of endocrine disruptors on steroidogenesis, which is the synthesis pathway of sex hormones, a validated in vitro method specified in the OECD TG-456 test guide was applied in our laboratory and ELISA and LC MS/MS were used to determine estradiol (E2) and testosterone (T) levels as a result of the experiment. It is aimed to use the methods of analysis and to compare the data obtained by both analysis methods. Partial validation of the method(s) was made using forskolin as a positive control (enzyme inducing effect) as stated in the OECD guideline, prochlorase as a negative control (enzyme inhibitory effect), ketoconazole, letrozole and genistein, which are reference substances with known efficacy on E2 and T in the steroidogenesis cycle. measurement performances were compared. Whether there is a difference between the two methods was evaluated statistically. Subsequently, 2-Methyl-1-H-indole-3-carboxyaldehyde (4-chlorophenyl) hydrazone (M6) and 2-methylindole-3-carboxyaldehyde, which are newly synthesized drug active substance candidates, will be recommended to be used as an aromatase enzyme inhibitor in the treatment of hormone-dependent breast cancer. The effects of phenyl hydrazone (M20) and their model compound, melatonin, on the entire steroidogenesis pathway were also investigated after the application of the H295R steroidogenesis method, by performing hormone analyzes with both methods. As a result, the two methods applied were evaluated in terms of advantages and disadvantages.
Endocrine disruptors (EB) are defined as exogenous substances that cause adverse health effects and changes in endocrine functions in healthy organisms or in their future generations. EB can act as an agonist or antagonist by interacting with hormone receptors, or it can also exert its effects by changing physiological hormone levels by interacting with enzymes involved in the biosynthesis and metabolism pathways of steroid hormones. In this thesis study, in order to investigate the effects of endocrine disruptors on steroidogenesis, which is the synthesis pathway of sex hormones, a validated in vitro method specified in the OECD TG-456 test guide was applied in our laboratory and ELISA and LC MS/MS were used to determine estradiol (E2) and testosterone (T) levels as a result of the experiment. It is aimed to use the methods of analysis and to compare the data obtained by both analysis methods. Partial validation of the method(s) was made using forskolin as a positive control (enzyme inducing effect) as stated in the OECD guideline, prochlorase as a negative control (enzyme inhibitory effect), ketoconazole, letrozole and genistein, which are reference substances with known efficacy on E2 and T in the steroidogenesis cycle. measurement performances were compared. Whether there is a difference between the two methods was evaluated statistically. Subsequently, 2-Methyl-1-H-indole-3-carboxyaldehyde (4-chlorophenyl) hydrazone (M6) and 2-methylindole-3-carboxyaldehyde, which are newly synthesized drug active substance candidates, will be recommended to be used as an aromatase enzyme inhibitor in the treatment of hormone-dependent breast cancer. The effects of phenyl hydrazone (M20) and their model compound, melatonin, on the entire steroidogenesis pathway were also investigated after the application of the H295R steroidogenesis method, by performing hormone analyzes with both methods. As a result, the two methods applied were evaluated in terms of advantages and disadvantages.
Açıklama
Anahtar Kelimeler
H295R, Steroidojenesis, Endokrin Bozucular, ELISA, LC MS/MS, H295R, Steroidogenesis, Endocrine Disruptors, ELISA, LC MS/MS
