Asitretin-Metotreksat Uygulanmış Ratların İnce Bağırsak Dokusunda Polifenol Oksidaz Aktivitesi Üzerine Alfa-Lipoik Asitin Etkileri

Yıl 2022, Cilt: 1 Sayı: 3, 28 - 37, 30.12.2022

Fatma Gönül Solmaz Emine İncilay Torunoğlu Emine Dıraman

Öz

Amaç: Asitretin, lipofilik zayıf bir asit olan suda az çözünen ve yağ dokusunda az biriken A vitamini analoğudur. Aynı zamanda anti-inflamatuar ve anti-proliferatif etkili non-immünosupresifdir. Timidilat sentetaz ve dihidrofolat redüktaz inhibitörü olan metotreksat ise bir folik asit analoğudur. Günümüzde de kanser tedavilerinin yanında mikozis fungoides, pitriyazis rubra pilaris, dermatomiyozit, sarkoidoz, egzama gibi hastalıkların tedavisinde de kullanılan Metotreksat aynı zamanda bir anti-folat antimetabolit ajandır. Ayrıca Asitretin ile kombinasyon halinde tedavide de kullanılmaya başlanmıştır. Bu çalışmada, Asitretin-Metotreksat uygulaması sonucunda meydana gelebilecek serbest radikallerin neden olduğu hasarın giderilmesinde güçlü bir antioksidan olan Alfa Lipoik Asit ratlara verilmiştir. Böylece oluşabilecek oksidatif hasarı onarma da önemli rol oynayan Alfa Lipoik Asitin ince barsak dokusundaki polifenol oksidaz (1.10.3.1) enzim aktivitesi üzerine etkisinin araştırılması amaçlanmıştır. Aktif bölgelesinde bakır içeren Polifenol oksidaz, fenolik bileşiklerin moleküler oksijen ile oksidasyonunu katalizleyen oksidoredüktaz sınıfı bir enzimdir.

Yöntem: Kontrol grubu, Alfa Lipoik Asit grubu, Asitretin+Metotreksat grubu ve Asitretin+Metotreksat+Alfa Lipoik Asit grubu olarak dört çalışma grubu oluşturulmuştur. Ratlara enjeksiyon işlemleri her sabah aynı saatte yapılmıştır ve enjeksiyondan 24 saat önce de aç bırakılmışlardır. Asitretin, Metotreksat ve Alfa Lipoik Asit % 0.9’luk NaCl’de çözülmüştür ve intraperitonal enjeksiyonla ratlara verilmiştir. Ratlar, enjeksiyondan sonraki 7. günde servikal dislokasyon ile sakrifiye edilmiştir. Bunu takiben kalp perfüzyonu işlemi yapılmış ve ince bağırsakları çıkarılmıştır. Örnekler önce homojenize edildi sonra sonikasyon ve santrifüj işlemleri uygulandı. Santrifüj işleminden sonra, elde edilen ince bağırsak homojenatları Polifenol oksidaz aktivite tayini için kullanılmıştır.

Bulgular ve Sonuç: Alfa Lipoik Asit, Asitretin + Metotreksat ve Asitretin + Metotreksat + Alfa Lipoik Asit grupları kontrol grubu ile karşılaştırılmıştır. Bunun sonucunda Alfa Lipoik Asit grubunun Kontrol grubu ile hemen hemen aynı aktiviteye sahip olduğu (%1,65 aktivasyon) gözlenirken Asitretin + Metotreksat grubu %10, Asitretin + Metotreksat + Alfa Lipoik Asit grubunun ise %25 aktivasyon gösterdiği gözlenmiştir. Asitretin + Metotreksat grubu ile Asitretin + Metotreksat + Alfa Lipoik Asit grubu karşılaştırıldığında Asitretin + Metotreksat + Alfa Lipoik Asit grubunun %13,5 daha fazla aktivasyon gösterdiği de belirlenmiştir. Alfa Lipoik Asit, tek başına verildiğinde ince bağırsak dokusunda Polifenol oksidaz aktivitesinde anlamlı bir fark göstermezken, Asitretin + Metotreksat kombinasyonu ile verildiğinde Polifenol oksidaz aktivitesini artırmıştır.

Anahtar Kelimeler

İnce bağırsak, asitretin, metotreksat, polifenol oksidaz, Small intestine, acitretin, methotrexate, polyphenol oxidase

Destekleyen Kurum

ONDOKUZ MAYIS ÜNİVERSİTESİ

Proje Numarası

PYO.FEN.1904.18.014

Effects of Alpha-Lipoic Acid On Polyphenol Oxidase Activity in Small Intestine Tissue of Rats Treated With Acitretin - Methotrexate Combination

Öz

Background and Aim: Acitretin is a vitamin A analogue, which is a lipophilic weak acid, less soluble in water and less accumulated in adipose tissue. It is also non-immunosuppressive with anti-inflammatory and anti-proliferative effects. Methotrexate, an inhibitor of thymidylate synthetase and dihydrofolate reductase, is a folic acid analogue. Today, Methotrexate, which is used in the treatment of diseases such as mycosis fungoides, pityriasis rubra pilaris, dermatomyositis, sarcoidosis, eczema, as well as cancer treatments, is also an anti-folate antimetabolite agent. It has also been used together with Acitretin in treatment. In this study, Alpha Lipoic Acid, which is a powerful antioxidant, was given to rats to eliminate the damage caused by free radicals that may occur as a result of Acitretin-Methotrexate application. Thus, it is aimed to investigate the effect of Alpha Lipoic Acid, which plays an important role in repairing the oxidative damage that may occur, on the polyphenol oxidase (1.10.3.1) enzyme activity in the small intestine tissue. Polyphenol oxidase, containing copper in its active site, is an oxidoreductase class enzyme that catalyzes the oxidation of phenolic compounds with molecular oxygen.

Materials and Methods: Four study groups were formed as Control Group, Alpha Lipoic Acid Group, Acitretin+Methotrexate Group and Acitretin+Methotrexate+Alpha Lipoic Acid Group. The injection procedures applied to the rats were performed at the same time every morning and they were fasted 24 hours before the injection. Acitretin, Methotrexate and Alpha lipoic acid were dissolved in 0.9% NaCl and given to rats by intraperitoneal injection. Rats were sacrificed by cervical dislocation on the 7th day after injection. Following this, cardiac perfusion was performed and the small intestines were removed. The samples were first homogenized, then sonication and centrifugation processes were applied. After centrifugation, the obtained small intestine homogenates were used for Polyphenol oxidase activity determination.

Result and Conclusion: Alpha Lipoic Acid, Acitretin + Methotrexate and Acitretin + Methotrexate + Alpha Lipoic Acid groups were compared with the control group. As a result, while it was observed that the Alpha Lipoic Acid group had almost the same activity with the Control group (1.65% activation), the Acitretin + Methotrexate group showed 10% activation, the Acitretin + Methotrexate + Alpha Lipoic Acid group showed 25% activation. When the Acitretin + Methotrexate group was compared with the Acitretin + Methotrexate + Alpha Lipoic Acid group, it was also determined that the Acitretin + Methotrexate + Alpha Lipoic Acid group showed 13.5% more activation. While Alpha Lipoic Acid did not show a significant difference in Polyphenol oxidase activity in small intestine tissue when given alone, it increased Polyphenol oxidase activity when given with Acitretin + Methotrexate combination.

Anahtar Kelimeler

Small intestine, acitretin, methotrexate, polyphenol oxidase

Proje Numarası

PYO.FEN.1904.18.014

Kaynakça

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