Boldine Exerts Neuroprotective Effects Against Rotenone Toxicity in SH-SY5Y In Vitro Model

Abstract

Purpose: Rotenone induces Parkinson's disease-like neurotoxicity through mitochondrial impairment, oxidative stress, and apoptosis. We investigated the neuroprotective efficacy of boldine against rotenone-induced cytotoxicity in human SH-SY5Y neuroblastoma cells. Material and Methods: SH-SY5Y cells were pretreated with boldine (0.1–100 µM) for 2 h before exposure to 200 nM rotenone for 24 h. Cell viability, clonogenic capacity, active caspase-3 levels, and apoptotic nuclear morphology were evaluated using the MTS assay, clonogenic assay, ELISA, and Hoechst 33342 staining, respectively. Results: Rotenone (200 nM) significantly reduced viability to 32.3%, suppressed clonogenic potential (3.9%), and increased active caspase-3 levels (p<0.0001). Pretreatment with 0.1 μ"M" boldine markedly restored cell viability to 92.2% and clonogenic activity to 81.9%, while significantly reducing caspase-3 levels and nuclear apoptotic alterations (p<0.0001). Higher boldine concentrations (≥5 μ"M" ) exhibited antiproliferative effects. Conclusion: Boldine exhibits a concentration-dependent biphasic response against rotenone-induced neurotoxicity, with marked neuroprotection at low concentrations and antiproliferative effects at higher concentrations. The protective effect of 0.1 µM boldine was associated with attenuation of caspase-3 activation and apoptotic nuclear alterations.

Keywords

• Boldine
• Rotenone
• SH-SY5Y
• Neurotoxicity
• Neuroprotection
• Caspase-3

Supporting Institution

Ege University Scientific Research Fund.

Project Number

TYL-2021-22429

Ethical Statement

All procedures performed in this study were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or compareable ethical standards. The Ethics Exemption Statement document is attached.

Boldine Exerts Neuroprotective Effects Against Rotenone Toxicity in SH-SY5Y In Vitro Model

Abstract

Amaç: Rotenon, mitokondriyal işlev bozukluğu, oksidatif stres ve apoptoz yoluyla Parkinson hastalığı benzeri nörotoksisiteye neden olmaktadır. Bu çalışmada, boldinin insan SH-SY5Y nöroblastoma hücrelerinde rotenon kaynaklı sitotoksisiteye karşı nöroprotektif etkinliği araştırıldı. Gereç ve Yöntem: SH-SY5Y hücreleri, 200 nM rotenona 24 saat süreyle maruz bırakılmadan önce 2 saat boyunca boldin (0.1–100 µM) ile ön işleme tabi tutuldu. Hücre canlılığı, klonojenik kapasite, aktif kaspaz-3 düzeyleri ve apoptotik nükleer morfoloji sırasıyla MTS testi, klonojenik analiz, ELISA ve Hoechst 33342 boyaması kullanılarak değerlendirildi. Bulgular: Rotenon (200 nM), hücre canlılığını anlamlı olarak %32.3’e düşürdü, klonojenik potansiyeli azalttı (%3.9) ve aktif kaspaz-3 düzeylerini artırdı (p<0.0001). 0.1 µM boldin ile ön işlem, hücre canlılığını belirgin şekilde %92.2’ye ve klonojenik aktiviteyi %81.9’a yükseltirken, kaspaz-3 düzeylerini ve apoptotik nükleer değişiklikleri anlamlı olarak azalttı (p<0.0001). Daha yüksek boldin konsantrasyonları (≥5 µM) antiproliferatif etkiler gösterdi. Sonuç: Boldin, rotenon kaynaklı nörotoksisiteye karşı konsantrasyona bağlı bifazik bir yanıt göstermekte; düşük konsantrasyonlarda belirgin nöroprotektif, daha yüksek konsantrasyonlarda ise antiproliferatif etkiler sergilemektedir. 0.1 µM boldinin koruyucu etkisi, kaspaz-3 aktivasyonunun ve apoptotik nükleer değişikliklerin azalması ile ilişkili bulunmuştur.

Keywords

• Boldin
• Rotenon
• SH-SY5Y
• Nörotoksisite
• Nöroproteksiyon
• Kaspaz-3.

Supporting Institution

Ege Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü

Project Number

TYL-2021-22429

Ethical Statement

Bu çalışmada gerçekleştirilen tüm işlemler, kurumsal ve/veya ulusal araştırma komitesinin etik standartlarına ve 1964 Helsinki Bildirgesi ile daha sonraki değişikliklerine veya eşdeğer etik standartlara uygun olarak yürütülmüştür. Etik Kurul Muafiyet Beyanı (Ethics Exemption Statement) belgesi ekte sunulmuştur.

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