AZOXYSTROBİN’İN İNSAN NÖROBLASTOMA HÜCRE HATTI ÜZERİNDEKİ İN VİTRO TOKSİK ETKİLERİ

Abstract

Amaç: Çalışmalar, tarım ilaçlarına maruziyetin otizm, beyin yaşlanması ve nörodejeneratif hastalıklar gibi sağlık sorunları ile ilişkili olabileceğini bildirmektedir. Yaygın kullanılan tarım ilaçlarından biri de strobilurin türevi bir fungusit olan azoksistrobin (AZS)’dir. Araştırmalar, çeşitli dokularda AZS’nin toksik etkilerini gösterse de nörotoksik etkilerin altında yatan temel mekanizmalar halen tam olarak anlaşılamamıştır. Tarım ilaçlarına maruziyetin sebep olduğu sağlık sorunlarının oksidatif stres ve apoptoz ile ilişkili olabieceği bildirilmektedir. Bu nedenle, nörotoksisite değerlendirmelerinde yaygın kullanılan bir in vitro model olan insan nöroblastoma (SH-SY5Y) hücrelerinde AZS’nin oksidatif hasar oluşturma ve hücre ölümünü indükleme etkisinin değerlendirilmesi amaçlanmıştır. Gereç ve Yöntem: SH-SY5Y hücreleri 24 saat boyunca AZS’nin farklı konsantrasyonlarına maruz bırakılmıştır. MTT testi ile sitotoksisite değerlendirilirken spesifik floresan boya ile reaktif oksijen türlerinin (ROS) oluşumu ve FITC Annexin V testi ile de hücresel apoptoz seviyeleri incelenmiştir. Bulgular: AZS’nin IC50 değeri 44.87 μM olarak bulunmuştur. ROS (en az 1,5 kat; p≤0,05) ve apoptoz (en az 5 kat; p≤0,05) seviyeleri 6.25-25 μM AZS maruziyet aralığında doza bağlı artmıştır. Sonuç: Bulgularımız, AZS’nin neden olduğu nörotoksik etkilerin ROS üretimi ve ROS ile indüklenen apoptozun bir sonucu olabileceğini düşündürmektedir. Ancak, AZS’nin toksik etki potansiyelinin daha iyi anlaşılması, mesleki ve çevresel risk faktörlerinin değerlendirilmesi, AZS kaynaklı nörotoksisiteye neden olan temel mekanizmaların aydınlatılması için ileri araştırmalara ihtiyaç duyulduğu da göz ardı edilmemelidir.

Keywords

• Azoksistobin
• apoptoz
• oksidatif stres
• nörotoksisite

IN VITRO TOXIC EFFECTS OF AZOXYSTROBIN ON HUMAN NEUROBLASTOMA CELL LINE

Abstract

Objective: Exposure to agricultural chemicals is associated with health issues such as autism, brain aging, and neurodegenerative diseases. Azoxystrobin (AZS), a strobilurin-derived fungicide, is a commonly used chemical. Although studies have demonstrated that AZS induces toxic effects on various tissues, there is a lack of clear understanding of the mechanisms underlying its neurotoxic effects. The mechanisms linking pesticide exposure to these health outcomes might be oxidative stress and apoptosis. Therefore, we conducted this study to investigate oxidative stress and cell death induction in human neuroblastoma SH-SY5Y cells, a widely established in vitro model for neurotoxicity experiments, after exposure to AZS. Materials and Methods: SH-SY5Y cells were exposed to different concentrations of AZS for 24 h. Cytotoxicity was evaluated using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide assay, the induction of reactive oxygen species (ROS) was determined using a fluorescent dye, and cell apoptosis was detected using the FITC Annexin V assay. Results: The half-maximal inhibitory concentration value of AZS was 44.87 μM. The levels of ROS (at least 1.5-fold; p≤0.05) and apoptosis (at least 5-fold; p≤0.05) increased in a dose-dependent manner at AZS exposure concentrations of 6.25-25 μM. Conclusion: AZS-induced neurotoxic effects may be a consequence of ROS generation and ROS-induced apoptosis. However, the mechanisms underlying AZS-induced neurotoxicity should be evaluated by further studies to gain a deeper understanding of its toxic capacity and conduct evaluations of occupational and environmental risks.

Keywords

• Azoxystrobin
• apoptosis
• oxidative stress
• neurotoxicity

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