Schizosaccharomycespombe’deTetrakonazol Kaynaklı Programlı Hücre Ölümü

Abstract

Sistemik triazol bir fungisit olan tetrakonazol tarımda ve insan sağlığında potansiyel toksik etkiler göstermektedir. Bu yüzden, sitotoksik etkileri ve eşlik eden mekanizmaları açığa çıkarılmalıdır. Bu çalışmada, tek hücreli biyoloji ve toksikoloji modeli olarak S. pombe (ED666) kullanılmıştır. Hücreler standart medyumda büyütülmüş, muameleler 30 C’de ve 180 rpm hızda çalkalamalı olarak yapılmıştır. 1-10 mg/L tetrakonazol doz-bağımlı hücre ölümüne sebep olmuştur. Apoptoz DAPI ve AO/EB boyamasıyla görüntülenmiştir. Aşırı ROS üretimi ve mitokondriyel bozulma DCFDA/NBT deneyleri ve Rhodamin 123 boyamasıyla gösterilmiş, bu sonuçlar da süperoksitdismutazlar ve glutatyonperoksidaz ifadelerindeki artışlarla desteklenmiştir. Potansiyel apoptotik genlerden biri olan Cnx1’in apoptozla ilişkisi transkripsiyonundaki artışla gösterilirken, diğer iki potansiyel gen, Pca1 ve Aif1 tetrakonazolden etkilenmemiştir. Sonuç olarak, tetrakonazol kaynaklı apoptoz ile, ROS hasarı ve mitokondriyel düzensizliğin (Cnx1-yoluyla) aracılık etmiş olduğu mekanizmalar S. pombe’de açıklığa kavuşturulmuştur.

Keywords

• Schizosaccharomyces Pombe
• Tetrakonazol
• Apoptoz
• Cnx1
• Oksidatif Hasar

Tetraconazole-induced Programmed Cell Death in Schizosaccharomyces pombe

Abstract

Tetraconazole, a systemic triazole fungicide, shows potential toxic effects in agriculture and human health. Therefore, its cytotoxic effects and accompanying mechanisms should be unraveled. S. pombe (ED666) was used in this study, as a unicellular biology and toxicology model. Cells were grown on standard media and all treatments were done at 30 C and shaking at 180 rpm 1-10 mg/L tetraconazole induced a dose-dependent cell death. Apoptosis was monitored by DAPI ve AO/EB staining. Excessive ROS production and mitochondrial impairment were shown by DCFDA/NBT assays and Rhodamine 123 staining, which were supported by increased expressions of superoxide dismutases and glutathione peroxidase. Involvement of one of the potential apoptotic genes, Cnx1, in apoptosis was shown by increased transcription whereas two other potential genes, Pca1 and Aif1, were not affected by tetraconazole treatment. In conclusion, tetraconazole-induced cytotoxicity and underlying mechanisms which were mediated via ROS damage and mitochondrial dysregulation (Cnx1-driven) were clarified in S. pombe.

Keywords

• Schizosaccharomyces Pombe
• Tetraconazole
• Apoptosis
• Cnx1
• Oxidative Damage

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