Tıbbi Bitki Mespilus Germanica Meyve Ekstraktının Kemoterapötik Potansiyelinin Değerlendirilmesi: Hücre Ölüm Yolakları ve DNA Hasar Mekanizması

Yıl 2024, , 202 - 207, 28.11.2024

Veysel Yüksek , Gül Görmez

https://doi.org/10.36483/vanvetj.1556349

Öz

Bitki ekstarktları antikanser dahil olmak üzere çeşitli biyolojik aktiviteler içeren doğal kompleks bileşiklerin bir karışımıdır. Sentetik ilaçlara göre daha az yan etkiye sahip olmaları, bitki ekstraktlarını kanser tedavisinde önemli bir strateji haline getirmiştir. Bu çalışmanın amacı, Mespilus germanica (Muşmula) meyve ekstraktın kemoterapötik potansiyelinin araştırılmasıdır. Elde edilen ekstraktın bileşik içeriği HPLC ile tespit edildi. MTT canlılık testi yoluyla proliferatif konsantrasyon (PRO) ve hücrelerin yarısının çoğalmasını inhibe eden konsantrasyon (IC50) belirlendi. PRO ve IC50 konsantrasyonlar 48 saat boyunca A549 akciğer kanser hücresine uygulandı. Çalışma grupları kontrol, PRO ve IC50 olmak üzere 3 grup olarak belirlendi. Elde edilen hücrelerden Trizol reagent-kloroform yöntemi kullanılarak toplam mRNA elde edildi. Toplam mRNA'dan cDNA sentezi yapıldı. Programlanmış hücre ölümü belirteçlerinin mRNA gen ekspresyon seviyeleri RT-qPCR ile tespit edildi. Tüm grup çalışmaları için p<0.05 istatistiksel olarak anlamlı kabul edildi. Ekstrakt içeriği analizinde klorojenik asit, elajik asit, kuersetin ve gallik asit polifenolik birleşikler tespit edildi. MTT sonucu IC50 540 μg/ml ve PRO 100 μg/ml olarak tespit edildi. IC50 konsantrasyonunun, ATG5 (otofajik) ve RIPK1 (nekrotik) genlerin ekspresyonunu önemli ölçüde arttırdığı (p<0.05) ortaya konuldu. Ayrıca Kaspaz-8, BAX, Apaf-1, Kaspaz-9, Kaspaz-3 ve Kaspaz-7 proteazlarının yanı sıra, genotoksik hasarla ilişkili genler olan PARP-1 ve P53'ün ekspresyonunun da önemli ölçüde arttığı gözlendi (p<0.05). Sonuç olarak Mespilus germenica A549 kanser hücrelerinin ortadan kaldırılması için istenilen programlanmış hücre ölüm yollaklarını tetiklediği tespit edildi. Mespilus germenica meyvesinin yeterli oranda tüketilmesinin kanser hücre çoğalmasını azaltabileceğini veya tamamen ortadan kaldırabileceği kanaatine varıldı. Elde edilen bu sonuçların kesin olarak ortaya konması için in vivo fazlı deneysel bir uygulamanın yapılması gerekeceği düşünülmektedir.

Anahtar Kelimeler

DNA hasarı, Hücre ölüm yolağı, Kanser, Mespilus Germanica

Evaluation of the Chemotherapeutic Potential of Medicinal Plant Mespilus germanica Fruit Extract: Cell Death Pathways and DNA Damage Mechanism

Öz

Plant extracts are a mixture of natural complex compounds containing various biological activities, including anticancer properties. The fact that they have fewer side effects than synthetic drugs has made plant extracts an important strategy in cancer treatment The purpose of this study was to explore the chemotherapeutic potential of Mespilus germanica (medlar) fruit extract. The compound content of the extract was determined by HPLC. The proliferative concentration (PRO) and the concentration inhibiting the proliferation of half of the cells (IC50) were determined by the MTT viability test. PRO and IC50 concentrations were treated to A549 lung cancer cells for 48 hours. The study groups were determined as 3 groups: control, PRO, and IC50. Total mRNA was obtained from the cells by using the Trizol Reagent-chloroform method. cDNA synthesis was performed from total mRNA. mRNA gene expression levels of programmed cell death markers were detected by RT-qPCR. For all group studies, p<0.05 was considered statistically significant. It was detected in the extract content analysis, chlorogenic acid, ellagic acid, quercetin, and gallic acid polyphenolic compounds. As a result of MTT, IC50 was detected at 540 μg/ml, and PRO was 100 μg/ml. It was revealed that IC50 concentration significantly increased (p<0.05) the expression of ATG5 (autophagic) and RIPK1 (necrotic) genes. In addition, it was observed that the expression of proteases Caspase-8, BAX, Apaf-1, Caspase-9, Caspase-3, and Caspase-7, as well as genes associated with genotoxic damage, PARP-1 and P53, increased significantly (p<0.05). As a result, it was determined that Mespilus germanica triggered the programmed cell death pathways in the A549 cancer cell line. It was concluded that adequate consumption of Mespilus germanica fruit can reduce or inhibit cancer cell proliferation. An experimental administration with an in vivo phase should be administered to reveal these results definitively.

Anahtar Kelimeler

Cancer, Cell death pathway, DNA damage, Mespilus germanica

Etik Beyan

Bu çalışma ticari olarak satın alınan hücre serisi üzerinde gerçekleştirildi. Etik onay gerektirmeyen belge sisteme yüklenmiştir.

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