Effect of Glucagon like Peptide-1 Analogue Exenatide on Neuron Survival in Rotenone-Induced In Vitro Parkinson's Disease Model

Abstract

Parkinson's disease (PD) is one the most common neurodegenerative disorder characterized by bradykinesia, tremor, rigidity and postural instability as a result of progressive degeneration of the dopaminergic neurons in substantia nigra pars compacta (SNc) and dopamine decrease in the striatum. Accumulating data recommends that combination of genetic and environmental factors including aging, exposure to pesticides, and heavy metals may have significant roles in the pathogenesis of PD. Rotenone, an organic pesticide, is commonly used for investigating the mechanism of neuronal degeneration in experimental PD models. In the present study, we aimed to investigate the effects of rotenone on SH-SY5Y human neuroblastoma cells, and also the neuroprotective effects of exenatide, a GLP-1 analogue, against rotenone-induced neurotoxicity. Briefly, to induce rotenone-induced neurotoxicity, SH-SY5Y cells were exposed to various doses of rotenone for 24 hours. To explore the neuroprotective effects of exenatide, different doses of exenatide were added into the medium with or without rotenone for 24 hours. Cell viability, clonegenic potential, morphological alterations, and apoptotic cell death were evaluated. Our results demonstrated a dose-dependent and significant decrease in cell viability in rotenone-exposed groups. Also, rotenone-treated group showed significant morphological alterations, decreased clonogenic potential and increased apoptosis. However, treatment of the cells with exenatide significantly improved cell viability and clonogenic activity, and reduced apoptotic cell death. In conclusion, these findings suggested that rotenone can induce neurotoxicity in neuronal cells and can be used to establish in vitro PD model. In addition, exenatide may have neuroprotective effects on neuronal cells against rotenone-induced neurotoxicity.

Keywords

• Parkinson,Rotenone,Exenatide,Neurotoxicity,Neuroprotective

Rotenon ile İndüklenen İn Vitro Parkinson Hastalığı Modelinde Glukagon Benzeri Peptid-1 Analoğu Ekzenatidin Nöron Sağkalımına Etkisi

Abstract

Parkinson hastalığı (PH), beyinde substansiya nigrada bulunan dopaminerjik nöronların hasarı ve buna eşlik eden striatumda dopamin eksikliği sonucunda gelişen, bradikinezi, rijidite ve postural instabilite ile seyreden kronik ve ilerleyici bir nörodejeneratif hastalıktır. Çalışmalar genetik ve yaşlanma, pestisid ve ağır metallere maruziyet gibi çevresel faktörlerin hastalığın etiyopatogenezinde önemli role sahip olabileceğine işaret etmektedir. Rotenon, deneysel PH modeli oluşturmak ve hastalığın mekanizmasını araştırmak amacıyla kullanılan organik bir pestisidtir. Bu çalışmada amacımız insan nöroblatoma hücre serisi olan SH-SY5Y hücrelerinde rotenonun toksik etkisini ve aynı zamanda GLP-1 analoğu olan ekzenatidin rotenon nörotoksisitesine karşı koruyucu etkili olup olmadığını araştırmaktır. Bu amaçla SH-SY5Y hücre kültürleri çeşitli dozlardaki rotenon ile 24 saat boyunca inkübe edildi. Ekzenatidin koruyucu etkisi yine farklı dozlardaki ekzenatidin rotenon ile birlikte kültür medyumuna eklenmesi ve 24 saat inkübe edilmesi ile incelendi. Rotenon ve ekzenatidin etkileri hücre canlılığı, klonojenik aktivite, morfolojik inceleme ve apopitotik hücre ölümünün değerlendirilmesi ile belirlendi. Sonuçlarımız rotenon eklenen gruplarda doza bağlı olarak ve anlamlı düzeyde hücre canlılığının ve klonojenik aktivitenin azaldığını gösterdi. Ayrıca rotenon eklenen hücrelerde belirgin olarak morfolojik değişim olduğu ve apopitotik hücre ölümünde artış olduğu gözlendi. Ortama ekzenatidin eklenmesi ise hücre canlılığının ve klonojenik aktivitenin korunmasına, apopitotik hücre ölümünde azalmaya neden oldu. Sonuç olarak; bulgularımız rotenonun SH-SY5Y hücrelerinde nörotoksik etkilerinin olduğunu ve in vitro PH modeli oluşturmak amacıyla kullanılabileceğini gösterdi. Ayrıca, sonuçlarımız ekzenatidin nöronal hücrelerde rotenon ile indüklenen toksisiteye karşı nöroprotektif etkiye sahip olabileceğini gösterdi.

Keywords

• Parkinson,Rotenon,Ekzenatid,Nörotoksisite,Nöroprotektif

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