Tenofovir alafenamide (TAF) içeren kitosan nanopartikülü formülasyonlarının hazırlanması, genotoksik ve mitotoksik etkilerinin araştırılması
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Dosyalar
Tarih
2020
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Attribution-NonCommercial-NoDerivs 3.0 United States
Attribution-NonCommercial-NoDerivs 3.0 United States
Özet
Amaç: TAF, 2016 yılında kronik hepatit B tedavisi için onaylanan, nükleotid revers transktiptaz inhibitör grubu (NRTI) yeni bir ilaçtır. Bu çalışmada, TAF ilacını içeren kitosan nanopartiküllerinin hazırlanması, karakterizasyonunun yapılması, TAF ve kombinasyonlarının mitotoksik/genotoksik etkilerinin araştırılması amaçlanmıştır. Materyal ve Metot: 45 µM TAF ve kitosan nanopartikülü, altı gün süresince HepG2 hücrelerine uygulanmıştır. TAF ve kombinasyonlarının olası genotoksik/sitotoksik etkileri; mikronükleus (Mn), komet ve apoptoz testleri ile değerlendirilmiştir. Mitotoksik etkiler ise; mtDNA, mitokondriyal membran potansiyeli (ΔΨm), süperoksit anyonları ile ROS seviyelerinin belirlenmesi ve RYR1, PCK1, ATF2 gen ifadelerinin değerlendirilmesi ile incelenmiştir. Bulgular: Genotoksik testler bakımından Mn ve komet değerleri; TAF ve TAF içeren kitosan nanopartikül gruplarında yüksek bulunmuştur, komet testi açısından ayrıca kitosan grubu da yüksek olarak belirlenmiştir (p<0.05). mtDNA seviyesinin göstergesi olarak COXII gen ifadesi değişiklikleri TAF grubunda düşük bulunurken; kitosan ve TAF içeren kitosan gruplarında yüksek bulunmuştur (p<0.05). ΔΨm için TAF ve TAF içeren kitosan uygulama gruplarının kontrole göre azaldığı görülmüştür (p<0.05). Hücresel ROS seviyesi, tüm uygulama gruplarında yüksek bulunmuştur (p<0.05). Annexin V ile apoptoz testinde, gruplar arasında farklılık görülmemiştir. Mitokondri-nükleus sinyal yolağı ile ilişkilendirilen genlerden; ATF2'nin gen ifade seviyesi değişmemiştir. RYR1 gen ifadesinin kitosan ve TAF gruplarında azaldığı, TAF içeren kitosan grubunda ise arttığı görülmüştür. PCK1 gen ifadesinin ise; TAF grubunda arttığı, kitosan ve TAF içeren kitosan gruplarında azaldığı görülmüştür (p<0.05). Sonuç: TAF'ın çalışılan dozda HepG2 hücrelerinde genotoksik ve mitotoksik etkileri olduğu görülmüştür. Kitosan ve kitosan ile enkapsüle TAF'ın, birçok parametrede genotoksik ve mitotoksik etkisi bulunmamıştır. Anahtar Kelimeler: Genotoksisite, Hepatit B, HepG2, kitosan enkapsülasyonu, mitotoksisite, TAF.
Aim: TAF is a new nucleotide reverse transcriptase inhibitor group (NRTI) drug, approved for the treatment of chronic hepatitis B in 2016. In this study, it was aimed to prepare, characterize chitosan nanoparticle containing TAF drug and to investigate mitotoxic/genotoxic effects of TAF drug and its combinations. Material and Method: 45 µM TAF and chitosan nanoparticle were applied to HepG2 cells for six days. Possible genotoxic/cytotoxic effects of TAF and its combinations were evaluated by (Micronuclei) Mn, comet and apoptosis tests. Toxic effects on mitochondria were investigated by evaluating mtDNA, mitochondrial membrane potential (ΔΨm), superoxide anions, ROS levels, and RYR1, PCK1, ATF2 genes expression. Results: In terms of genotoxic tests, Mn and comet values were found higher in TAF and and chitosan nanoparticle containing TAF groups, and chitosan group was also determined for comet test (p<0.05). As an indicator of mtDNA level, COXII gene expression changes were found low in the TAF group, whereas chitosan and chitosan containing TAF groups were high (p<0.05). For ΔΨm, it was observed that chitosan containing TAF and TAF application groups decreased compared to the control (p<0.05). Cellular ROS level was found higher than control in all application groups (p<0.05). There was no difference between the groups in apoptosis test with Annexin V (p>0.05). Among the genes associated with the mitochondria-nucleus signal pathway, the level of gene expression of ATF2 has not changed. RYR1 gene expression increased in chitosan containing TAF group while decreased in chitosan and TAF groups. It was observed that PCK1 gene expression increased in TAF group and decreased in chitosan and chitosan containing TAF groups (p<0.05). Conclusion: TAF has been shown to have genotoxic and mitotoxic effects in HepG2 cells at the studied dose. TAF encapsulated with chitosan and chitosan has not been found to have genotoxic and mitotoxic effects in many parameters. Key Words: Chitosan encapsulation, genotoxicity, hepatitis B, HepG2, mitotoxicity, TAF.
Aim: TAF is a new nucleotide reverse transcriptase inhibitor group (NRTI) drug, approved for the treatment of chronic hepatitis B in 2016. In this study, it was aimed to prepare, characterize chitosan nanoparticle containing TAF drug and to investigate mitotoxic/genotoxic effects of TAF drug and its combinations. Material and Method: 45 µM TAF and chitosan nanoparticle were applied to HepG2 cells for six days. Possible genotoxic/cytotoxic effects of TAF and its combinations were evaluated by (Micronuclei) Mn, comet and apoptosis tests. Toxic effects on mitochondria were investigated by evaluating mtDNA, mitochondrial membrane potential (ΔΨm), superoxide anions, ROS levels, and RYR1, PCK1, ATF2 genes expression. Results: In terms of genotoxic tests, Mn and comet values were found higher in TAF and and chitosan nanoparticle containing TAF groups, and chitosan group was also determined for comet test (p<0.05). As an indicator of mtDNA level, COXII gene expression changes were found low in the TAF group, whereas chitosan and chitosan containing TAF groups were high (p<0.05). For ΔΨm, it was observed that chitosan containing TAF and TAF application groups decreased compared to the control (p<0.05). Cellular ROS level was found higher than control in all application groups (p<0.05). There was no difference between the groups in apoptosis test with Annexin V (p>0.05). Among the genes associated with the mitochondria-nucleus signal pathway, the level of gene expression of ATF2 has not changed. RYR1 gene expression increased in chitosan containing TAF group while decreased in chitosan and TAF groups. It was observed that PCK1 gene expression increased in TAF group and decreased in chitosan and chitosan containing TAF groups (p<0.05). Conclusion: TAF has been shown to have genotoxic and mitotoxic effects in HepG2 cells at the studied dose. TAF encapsulated with chitosan and chitosan has not been found to have genotoxic and mitotoxic effects in many parameters. Key Words: Chitosan encapsulation, genotoxicity, hepatitis B, HepG2, mitotoxicity, TAF.
Açıklama
Anahtar Kelimeler
Genetik, Genetics
Kaynak
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Scopus Q Değeri
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Künye
Sezer, Selcen (2020). Tenofovir alafenamide (TAF) içeren kitosan nanopartikülü formülasyonlarının hazırlanması, genotoksik ve mitotoksik etkilerinin araştırılması. Yayımlanmış Doktora tezi, İnönü Üniversitesi, Malatya.1-122 ss.
