Overexpression Of PGC-1α In The Protective Effect Of The Degeneration Of Dopaminergic Neurons

BackgroundThe Parkinson’s disease (PD) is a nervous system retrogression disease whichcommonly happens in the elderly. Though there are many facters that could triggerthe PD, such as the environment and the gene elements, the mitochondrial dysfunctionis one of the common ground. Therefore, rapid control the function of mitochondrialhas become a potential therapeutic target. In recent years’ studies, it’s researched thatthe Peroxisome Proliferator-Activated Receptor γ (PPARγ) co-activator1α (PGC-1α)takes a prominent role in the process of mitochondrial biological overexpression andoxidative stress. However, there is no report about the using of PGC-1as controllingthe mitochondrial function, to avoid enducing the SH-SY5Y cells’ oxidative pressdegeneration of dopaminergic neurons caused by the MPP~+.ObjectiveTo research the PGC-1α overexpression protection SH-SY5Y cells fromapoptosising caused by MPP~+, and analyzing it’s molecular mechanisms.Methods1. The MTT method was used to detect the cell’s survival rate. Then we observe thevitality changes of the SH-SY5Y cell which infectedby different amount viruses after48hours to choose the fittest virus infection coefficient and infection time. And weobserve the vitality changes of the SH-SY5Y cell which infected by different MPP~+concentration after24hours to choose the best MPP~+concentration. And we observethe vitality changes of SH-SY5Y cell separately treated by the Con, MPP~+,thevacancy viruses Ad+Con, the vacancy viruses Ad+MPP~+, the Ad-GFP-PGC-1α+Con,and the Ad-GFP-PGC-1α+MPP~+.2. The flow cytometryanalysis technology has used to detect the ROS level and themitochondrial membrane potential level inside the SH-SY5Y cell.3. The multifunctional enzyme mark instrument of fluorescence method has used to measure the amount of H2O2emerged inside the SH-SY5Y cell. And thechemiluminescence method has used to measure the ATP level inside the cell.4. The Real-time RCR can be semi-quantitative for the expression of PGC-1α andNRF-1mRNA inside of the SH-SY5Y cell.Results1.MTT method determination found that cell viability decreased slightly when treatedby MPP~+, and it doesn’t have significant change compared with blank when treated byPGC-1α, and it significant decreased when treated by Ad+MPP~+. However, the cellviability is significantly higher then Ad+MPP~+treatment when treated byAd-GFP-PGC-1α+MPP~+.2.Flow cytometry detection the ROS levels showed that the PGC-1α can inhibit MPP~+inducing SH-SY5Y cells’ ROS generation. And Multifunctional microplate readerfluorescence method detected that the PGC-1α can inhibit MPP~+inducing SH-SY5Ycells’ generation H2O2and ROS. And Flow cytometry detection showed that themitochondrial membrane potential increased when using PGC-1α. And themultifunction microplate reader chemical strength detected that PGC-1α can improvethe level of ATP.3.Real-time RCR result showed that the cell treated by MPP~+PGC-1α increased(P>0.05), and NRF1mRNA increase (P<0.05). But the PGC-1α and NRF1mRNAlevel decrease in cell when treat by transfected PGC-1α adenovirus.Conclusions1. In this experiment, the cellular damaged by MPP~+was used as the model for PD,which is a traditional PD external model. The molding concentrations have somedifference in which the models were used, but it’s nearly the same. The result may beassociated with the choosed cell line and the cell culture medium. Therefore, we canchoose the appropriate modeling concentration for this experiment determined byMTT method in the different experiment conditions.2. The overexpression of PGC-1can decrease the production of ROS and H2O2,increase the mitochondrial membrane potential and ATP level.That’s why the PGC-1can decrease the injury introduced by MPP~+oxidative stress. 3. The increase of PGC-1increased the expression of NRF1gene. According to thisphenomenon, we can conjecture that the PGC-1decrease the injury of oxidativestress by NRF1path, rather than involving in oxidative stress directly.4. Based on previous studies and our experiment, it’s found that the PGC-1may beaideal target for slowing down the defection rate of mitochondrial function inneurodegenerative disease.