In vitro parkinson hastalığı modelinde timokinonun apoptotik mekanizmalar üzerine etkisi
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Dosyalar
Tarih
2019
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi, Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Amaç: Parkinson Hastalığı (PH) ciddi yaşam konforu ve iş gücü kaybına hatta ölüme sebep olan, toplumsal olarak da ülke ve dünya ekonomisine ciddi bir yük oluşturan nörodejeneratif bir hastalıktır. Yaşla birlikte ilerleyen bu hastalık, insan ömrünün uzadığı günümüzde daha da fazla önem kazanmıştır. Ancak henüz kesin bir tedavi, veya koruyucu bir yöntem geliştirilememiştir. Biz bu çalışmada PH'de timokinonun koruyucu bir ajan olarak potansiyelini inceledik. Gereç ve Yöntem: Çalışmamızda SH-SY5Y hücrelerine 6-hidroksidopamin (6-OHDA) uygulamasıyla oluşturulmuş PH hücre kültürü modeli kullanılmıştır. Modele uygulanacak 6-OHDA ve timokinonun doz ve uygulama süresinin belirlenmesi ve timokinonun nöron koruyucu etkisinin gösterilmesi için MTT ile spektrofotometrik inceleme yapılarak hücre canlılığı tespit edilmiştir. 6-OHDA 100 μM ve timokinon 2.5 μM konsantrasyonda uygulanmıştır. Western Blot yöntemiyle 6-OHDA'nın Bax proteini ve 6-OHDA ve timokinonun kaspaz-3, bcl-2 ve bcl-xl protein düzeylerine etkisi incelenmiştir. Elde edilen veriler One way ANOVA ve Tukey testleri ile analiz edilmiştir. Bulgular: MTT testinde timokinonun hücre canlılığını arttırdığı, nöronlarda koruyucu bir etkisi olduğu görülmüştür (p<0.05). 6-OHDA, bax'ı 6 ve 12 saat 100 μM konsantrasyonda uygulandığında arttırmıştır (p<0.05). Timokinon ve 6-OHDA, Bcl-2'yi 6 saat uygulamada etkilememiştir (p>0.05), 12 saatte ise 6-OHDA Bcl-2'yi kontrole göre azaltmış (p<0.05), timokinon + 6-OHDA ise 6-OHDA uygulamasına göre Bcl-2'yi arttırmıştır (p<0.0005). 6 saat uygulamada 6-OHDA bcl-xl düzeyini azaltmış (p<0.005), timokinon + 6-OHDA, 6-OHDA uygulamasına göre bcl-xl düzeyini arttıramamıştır (p>0.05), ancak timokinon tek başına uygulandığında bcl-xl düzeyini arttırmıştır (p<0.05). Timokinon 6 ve 12 saat uygulamada kaspaz-3 düzeyini arttırırken (p<0.0005), 6-OHDA azaltmıştır (p<0.0005). 6 saat uygulanan timokinon + 6-OHDA, 6-OHDA uygulamasına göre kaspaz-3'ü arttırmıştır (p<0.0005) ancak 12 saat uygulamada bu iki değer arasında anlamlı bir fark bulunmamıştır (p>0.05). Sonuç: Bu çalışma verileri sonucunda timokinonun SH-SY5Y hücre canlılığını koruduğu, bcl-2 ve bcl-xl düzeylerini de arttırdığı görülmüştür. Dolayısıyla timokinonun nöron koruyucu bir etkisinin olduğu ve bu etkiyi apoptoz mekanizmasını baskılayarak oluşturduğu gösterilmiştir. Bu bulgular ışığında timokinonun PH'de nöroprotektif etkilerinin olduğu düşünülmekle beraber, daha ileri çalışmalara ihtiyaç duyulmaktadır.
Objective: Parkinson’s Disease (PD) is a neurodegenerative disease that seriously hampers life standards and working ability and can even cause death. It is also a serious burden on the economy of our country and world. Human life span has greatly increased in recent years and so diseases that progresses with age like PD have become more important. Still, an effective cure or protection method against this disease could not yet be developed. In this study, we investigated thymoquinons potential as a protective agent. Methods: In our study we have used a PD model of 6-OHDA administrated SH-SY5Y cells. To determine 6-OHDA and thymoquinone doses we would use on our model and whether thymouinone has a neuroprotective effect, we have used spectrophotometer analysis with MTT cell viability test. We determined the doses of thymoquinone and 6-OHDA as 2.5 μM and 100 μM respectively. Western blotting method was used to determine the effect 6-OHDA had on the apoptotic protein Bax and 6-OHDA and thymoquinone had on the caspase-3, bcl-2 and bcl-2. Statistical analyisis was made using the Tukey and ANOVA tets. Results: MTT test showed us thymoquinone improved cell viability and so has a neuroprotective property (p<0.05). In the western blot test we saw 6-OHDA increased Bax levels when administrated for 6 and 12 hours (p<0.05). Thymoquinone and 6-OHDA did not have a significant effect on bcl-2 when administrated for 6 hours, but in 12 hours 6-OHDA decreased bcl-2 levels significantly compared to control (p<0.05) and thymoquinone increased bcl-2 compared to 6OHDA (p<0.05). In 6 hours adminitrastion, 6-OHDA decreased bcl-2 levels significantly (p<0.005) but thymoquinone had no significant effect on bcl-2 levels compared to 6-OHDA (p>0.05). However when given without 6-OHDA, thymoquinone could make significant increase in bcl-2 levels compared to control (p<0.05). Thymoquinone increased and 6-OHDA decreased caspase-3 levels significantly when administrated for both 6 and 12 hours (p<0.0005). In 6 hours, thymoquinone increased caspase-3 levels compared to 6-OHDA (p<0.0005) but showed no significant change in 12 hours (p>0.05). Conclusions: This study shows thymoquinone preserves cell viability of SH-SY5Y cells and increases bcl-2 and bcl-2 proteins. Thus it was deduced that thymoquinone has a neuroprotective property which takes effect through inhibiting apoptosis. In light of these findings, thymoquinone can be said to hold promise for a future protective PD medicine, however to accomplish this, further research needs to be cunducted.
Objective: Parkinson’s Disease (PD) is a neurodegenerative disease that seriously hampers life standards and working ability and can even cause death. It is also a serious burden on the economy of our country and world. Human life span has greatly increased in recent years and so diseases that progresses with age like PD have become more important. Still, an effective cure or protection method against this disease could not yet be developed. In this study, we investigated thymoquinons potential as a protective agent. Methods: In our study we have used a PD model of 6-OHDA administrated SH-SY5Y cells. To determine 6-OHDA and thymoquinone doses we would use on our model and whether thymouinone has a neuroprotective effect, we have used spectrophotometer analysis with MTT cell viability test. We determined the doses of thymoquinone and 6-OHDA as 2.5 μM and 100 μM respectively. Western blotting method was used to determine the effect 6-OHDA had on the apoptotic protein Bax and 6-OHDA and thymoquinone had on the caspase-3, bcl-2 and bcl-2. Statistical analyisis was made using the Tukey and ANOVA tets. Results: MTT test showed us thymoquinone improved cell viability and so has a neuroprotective property (p<0.05). In the western blot test we saw 6-OHDA increased Bax levels when administrated for 6 and 12 hours (p<0.05). Thymoquinone and 6-OHDA did not have a significant effect on bcl-2 when administrated for 6 hours, but in 12 hours 6-OHDA decreased bcl-2 levels significantly compared to control (p<0.05) and thymoquinone increased bcl-2 compared to 6OHDA (p<0.05). In 6 hours adminitrastion, 6-OHDA decreased bcl-2 levels significantly (p<0.005) but thymoquinone had no significant effect on bcl-2 levels compared to 6-OHDA (p>0.05). However when given without 6-OHDA, thymoquinone could make significant increase in bcl-2 levels compared to control (p<0.05). Thymoquinone increased and 6-OHDA decreased caspase-3 levels significantly when administrated for both 6 and 12 hours (p<0.0005). In 6 hours, thymoquinone increased caspase-3 levels compared to 6-OHDA (p<0.0005) but showed no significant change in 12 hours (p>0.05). Conclusions: This study shows thymoquinone preserves cell viability of SH-SY5Y cells and increases bcl-2 and bcl-2 proteins. Thus it was deduced that thymoquinone has a neuroprotective property which takes effect through inhibiting apoptosis. In light of these findings, thymoquinone can be said to hold promise for a future protective PD medicine, however to accomplish this, further research needs to be cunducted.
Açıklama
Anahtar Kelimeler
Parkinson Hastalığı, Timokinon, 6-Hidroksidopamin, SH-SY5Y, Parkinson’s Disease, Thymoquinone, 6-Hydroxydopamine
