Lazer aksotomi sonrası primer kortikal nöron kültüründe rifampisinin nöronal hücre sağkalımı üzerine etkisi

Year 2019, Volume: 3 Issue: 2, 135 - 140, 23.08.2019

Mehmet Ozansoy , Ebru Coşkun Muzaffer Beyza Çetin Ozansoy Şeyda Çankaya , Mehmet Yalçın Günal Zübeyir Bayraktaroğlu Burak Yuluğ , Lütfü Hanoğlu

https://doi.org/10.30565/medalanya.553846

Abstract

Amaç: Aksonal hasarlanma sonucu oluşan nörodejenerasyon travmatik beyin hasarı ve omurilik yaralanmalarında sıklıkla görülen bir tablodur. Sinaptik disintegrasyon nedeniyle trofik faktörlerin nöron gövdesine retrograd olarak taşınamaması sonucu trofik faktör yetersizliği aksonu hasarlanmış olan nöronun ölümüne yol açar. Rifampisin antibiyotik etkilerinin yanı sıra çeşitli nörodejeneratif süreçlerde nöroprotektif etkileri de olan bir antibiyotiktir. Bu çalışmadaki amacımız lazer aksotomi ile hasarlanmış primer kortikal nöronlarda rifampisinin akut nöroprotektif etkilerinin araştırılmasıdır. 

Yöntemler: Yenidoğan farelerin kortikal nöronları izole edilerek kültür edildi. Yirmi dört saat sonra seçilen üç rifampisin dozu (1 μM, 10 μM ve 100 μM) primer kortikal nöronlara uygulandı. Rifampisin uygulamasından 15 dakika sonra lazer aksotomi gerçekleştirildi. Aksotomiden 24 saat son ra propidiyum iyodit boyaması ile nöronların canlılığı ölçüldü.

Bulgular: Lazer aksotomi nöronal canlılığı %80,45 oranında azalmaktadır. Buna karşılık rifampisin ön uygulaması ise aksonları hasarlı nöronların canlılığını seçilen üç dozda da anlamlı ölçüde arttırmıştır. 

Sonuç: Lazer aksotomi ile hasarlanmış kortikal nöronlardaki canlılığı arttırması nedeniyle rifampisinin söz konusu deneysel modelde akut nöroprotektif etkisi olduğu düşünülmektedir. 

Keywords

Lazer aksotomi, rifampisin, nöronal canlılık, primer kortikal nöron, nörotravma

The effect of rifampicin on the neuronal cell survival in prımary cortical neuron culture after laser axotomy

Abstract

Aim: Neurodegeneration caused by the axonal injury is a widely seen phenomenon in spinal cord and traumatic brain injuries. Due to the disintegration of the synaptic connection neurotrophic factors could not be transported retrogradely towards the cell body and the deprivation of the trophic factors lead to the degeneration and death of the injured neuron. Rifampicin is an antibiotic exhibiting several neuroprotective functions in various neurodegenerative conditions. Here we aim to investigate the acute neuroprotective effect of rifampicin in primary cortical neuron culture in which neurons are axotomized by laser axotomy.

Methods: Neonatal male mice were used in order to isolate cortical neurons. Isolated primary cortical neurons were cultured. After 24 hours three different rifampicin concentrations (1 μM, 10 μM and 100 μM) were applied to the neurons and after 15 minutes of rifampicin addition, neurons were laser axotomized. Viability of the cells was evaluated by propidium iodide staining after 24 hours of axotomy. 

Results: Laser axotomy decreases the cortical neuron viability significantly by 80.45%, while rifampicin pre-treatment increases their viability in all three dosages in a statistically significant manner. 

Conclusion: Rifampicin has an acute neuroprotective effect on the viability of the laser axotomized cortical neurons. 

Keywords

Laser axotomy, rifampicin, neuronal viability, primary cortical neuron, neurotrauma

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