Onopordum acanthium Özütünün Acanthamoeba castellanii Trofozoitleri Üzerinde Amoebisidal Etkisinin, DNA Hasarına Karşı Koruyucu ve Sitotoksik Aktivitelerinin Araştırılması

Yıl 2025, Cilt: 15 Sayı: 1, 434 - 447, 15.03.2025

Bülent Kaynak , Gülizar Aydoğdu , Zeynep Kolören

https://doi.org/10.31466/kfbd.1586799

Öz

Çalışmada Onopordum acanthium’un etanolik kök özütünün; Acanthamoeba castellanii trofozoitlerine karşı amoebisidal aktivitesi ile özütün kimyasal kompozisyonu, memeli hücrelerinde sitotoksisitesi ve DNA koruyucu aktivitesi araştırılmıştır. Farklı konsantrasyonlardaki (82, 41, 20,5, 10,25, 5,125, 2,56, 1,28 mg/mL) özütün A. castellanii trofozoitleri üzerinde amoebisidal etkisi tespit edilmiştir. Özütün, epitel hücre olduğu için seçilen HeLa (serviks adenokarsinomu) hücre hattında sitotoksisitesi 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) analiziyle araştırılmıştır. Özütün fitobileşikleri Gaz Kromatografisi Kütle Spektrometresi (GC-MS) ile saptanmıştır. Hidroksil radikaliyle pBR322 plazmid DNA’sında oluşturulan DNA hasarına karşı özütlerin DNA koruyucu potansiyeli araştırılmıştır. O. acanthium özütünün 82 mg/mL konsantrasyonda 72. saatte canlı trofozoite rastlanmadığı gözlenmiştir. IC50 değerinin sırasıyla 72., 48., 24. saatlerde 3,09, 5,55 ve 13,6 mg/mL olarak tespit edilmiştir. MTT analizi sonucunda, özütün farklı konsantrasyonlarıyla (14, 13, 11, 9,8, 7, 4, 3, 1,50, 0,44, 0,22 mg/mL) muamele edilen HeLa hücrelerinin canlılığı doza bağlı olarak azalmıştır. 72. saatte IC50 değeri 8,04 mg/mL olarak tespit edilmiştir. Hidroksil radikali ile indüklenen DNA hasarını engelleme üzerine etkileri incelendiğinde, özütün 20,5, 10,25, 5,125, 2,56, 1,28 mg/mL konsantrasyonlarda DNA hasarını engelleme etkisinin olduğu tespit edilmiştir. Elde edilen verilerle özütün günümüzde Acanthamoeba enfeksiyonlarının tedavisinde kullanılan ilaçlara alternatif olarak önerilebilecek bir aday olduğu düşünülmektedir.

Anahtar Kelimeler

Acanthamoeba castellanii, GC-MS, HeLa Hücresi, Onopordum acanthium, pBR322 Plazmid DNA

Etik Beyan

Araştırma için etik Onay gerekmemektedir.

Investigation of Onopordum acanthium Extract Regarding Its Amoebicidal Activity Against Acanthamoeba castellanii Trophozoites, DNA-protecting and Cytotoxic Activities

Öz

The amoebicidal activity of the ethanolic root extract of Onopordum acanthium against Acanthamoeba castellanii trophozoites, its chemical composition, cytotoxicity in mammalian cells, and DNA protective activity were investigated. The amoebicidal effect of the extract was assessed at various concentrations (82, 41, 20,5, 10,25, 5,125, 2,56, 1,28 mg/mL) on A. castellanii trophozoites. The cytotoxicity of the extract in the HeLa (cervix adenocarcinoma) cell line, which was selected due to it is an epithelial nature, was evaluated using the 3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) analysis. The phyto-compounds present in the extract were identified using Gas Chromatography-Mass Spectrometry (GC-MS). The DNA protective potential of the extracts against hydroxyl radical induced DNA damage in pBR322 plasmid DNA was also examined. At a concentration of 82 mg/mL of O. acanthium extract, no viable trophozoites were observed at the 72nd hour. The IC50 values were 3,09, 5,55 and 13,6 mg/mL at 72, 48 and 24 hours, respectively. MTT analysis revealed a dose-dependent decrease in the viability of HeLa cells treated with various concentrations of the extract (14, 13, 11, 9,8, 7, 4, 3, 1,50, 0,44, 0,22 mg/mL), with an IC50 value of 8,04 mg/mL at 24 hours. Furthermore, the extract exhibited protective effects against DNA damage induced by hydroxyl radicals at concentrations of 20,5, 10,25, 5,125, 2,56 and 1,28 mg/mL. According to our results, the extract is proposed as a potential alternative to current medications used to treat Acanthamoeba infections.

Anahtar Kelimeler

Acanthamoeba castellanii, GC-MS, HeLa Cell, Onopordum acanthium, pBR322 Plasmid DNA

Kaynakça

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