Selegiline Inhibits 3-NP-induced Cell Apoptosis And Its Mechanism In Microglial Cells

Background:Parkinson's disease(PD)is a neurodegenerative disease characterized by the progressive degeneration of specific neurons in the brain of the affected individual.The pathogenesis of Parkinson's disease is still unknown.icroglia-mediated neuroinflammation,oxidative stress and mitochondrial dysfunction are thought to play important roles in the degeneration of dopamine neurons.Oxidative stress and mitochondrial dysfunction can directly affect energy metabolism and ultimately affect the role of abnormal protein phagocytosis in microglia cells,thus damaging dopaminergic neurons.Many studies have confirmed that silegiline has anti-oxidative stress,protects mitochondrial function,and reduces the accumulation of neuron-synaptic nuclear protein.However,there is no in vitro experiment on the protective effect of silegiline on microglia in nervous system.Objective:The effect of silegilan on the apoptosis of bv-2 cells in mitochondrial oxidative stress was preliminarily elucidated.Methods:1.Different concentrations of 3-NP(2 mM,3 mM,4 mM)were used to act BV-2 cells.MTT assay was used to detect the cell proliferation activity,calculate the cell growth inhibition rate,and select 3-NP with a cell growth inhibition rate of 50%Concentration established oxidative stress injury model.2.Selegiline acted on bv-2 cells alone,and the concentration gradients were set as 100 mm,150m,200 m,250m,300 m,400m,500 m,900m and 1000 m,respectively,to detect the effect of selegiline on the growth inhibition rate of BV2 cells within a certain concentration range.3.According to the co-culture of 3-np concentration and selegiline concentration screened out in the first two experiments,the intervention effect ofselegiline on 3-np damaged bv-2 cells was detected by MTT method.4.Immunohistochemistry was used to detect the expression of bcl-2,Bax,caspase-3 and caspase-9 in each group.5.The expression of Bcl-2,Bax,Caspase-3 and Caspase-9 in each group was detected by TUNEL method.6.Expression of Bcl-2,Bax,Caspase-3 and Caspase-9 in each group was de-tected by flow cytometry.Results:1.Toxicity of 3-NP to BV2 cells The 24 hMTT results showed that different concentrations of 3-NP can reduce the survival rate of BV2 cells in a concentration-dependent manner.When the 3-NP concentration was 3 mM,the cell growth inhibition rate was 47.50 ± 0.73%.2.The concentration of 100 ?M,150 ?M,200 ?M,250 ?M,300 ?M,400 ?M,500 ?M,900 ?M,and 1000 ?M selegiline hydrochloride solution was applied to BV2 cells alone.MTT results showed that: The growth inhibition rate of cells has no significant effect,and only when the concentration is greater than 900 ?M,selegiline can inhibit the growth of BV-2 cells.3.MTT results of selegiline hydrochloride at a concentration of 100 ?M and200 ?M on co-culture of 3-NP-damaged cells showed that selegiline hydrochloride could significantly interfere with the growth inhibitory effect of 3-NP on BV2 cells(P<0.05).4.Immunohistochemical results showed that 200 ?M selegiline significantly reduced the expression of Bax,Caspase-3,and Caspase-9 in BV-2 cells(P <0.01).5.The results of TUNEL showed that the final concentration of 3-NP 3 mM could lead to apoptosis of BV-2 cells,and 200 ?M of selegiline could effectively protect BV-2 cells(P <0.01).6.Flow cytometry results showed that 200 ?M of selegiline and 3-NP3 mM could reduce the apoptosis rate of BV2 cells.Conclusions:1.Mice were BV-2 cells were exposed to a concentration of injury model could induce oxidative stress microglia 3-NP solution of 3mM.2.Selegiline pretreatment can significantly reduce the activity and apoptosis of BV-2 cells after 3-NP injury.3.Selegiline can regulate the apoptosis induced by oxidative stress injury of BV-2 cells by 3-NP through mitochondrial pathway.