Effect of Thymoquinone on Autophagy-Associated Proteins in 6-Hydroxydopamine-Induced Neurodegeneration Model
Project no. 144 · Status: Kapanmış · 2018
Summary
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. 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. 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). 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. Key Words: Parkinson’s Disease; Thymoquinone; 6-Hydroxydopamine; SH-SY5Y