MA-5 Activates AMPK Signal Pathway To Enhance Mitochondrial Autophagy In The Treatment Of Parkinson’s Disease

【Objective】To investigate the mechanism of action of MA-5 in the treatment of Parkinson’s disease and to clarify whether MA-5 protects MPTP-induced PD mice by enhancing mitophagy through the AMPK signaling pathway.【Methods】1.Animal model establishment: MPTP(30mg/kg/d)was injected intraperitoneally for 7 consecutive days to construct a mouse model of Parkinson’s disease.2.Behavioral experiment : Observe the average speed and total distance of the mice’s movement through open field experiments,and record the movement trajectory of the mice,so as to evaluate the movement ability of the mice.3.Tyrosine hydroxylase detection : Western Blot detects the expression of TH protein.4.Inflammatory factor determination:The expression of TNF-α,IL-6 and IL-1 βwas detected by Western Blot.5.Analysis of mitophagy related proteins:The expression of LC3-II/LC3-I,p62,PINK1 and Parkin in brain tissue was evaluated by Western Blot.6.Pathway-related protein detection:The levels of p-AMPK and pm TOR were detected by WB.【Results】1.After continuous intraperitoneal injection of MPTP for 7 days,the open field test showed that the average speed and movement distance of mice treated with MPTP decreased,indicating that the PD mice were successfully modeled.Compared with the control group,the expression of TH in MPTP group decreased,while the levels of pro-inflammatory cytokines IL-6,IL-1β and TNF-α increased.The treatment of MA-5 can improve the motor ability of PD mice,upregulate the expression of TH protein,and reduce the expression of proinflammatory cytokines IL-6,IL-1 β and TNF-α in the brain tissue of PD mice.2.Treatment of mice with cyclosporine A,an inhibitor of mitophagy,reversed MA-5 effects in PD mice.Compared with MA-5 + MPTP group,the mean speed and distance of PD mice were decreased,TH protein expression and mitophagy-related indicators LC3-II/LC3-I,PINK,and Parkin expression were decreased,while p62 expression was increased,and the levels of proinflammatory cytokines IL-6,IL-1β,and TNF-α in brain tissue were increased after pretreatment with autophagy inhibitor CSA.3.With the intervention of AMPK inhibitor Compound C,the protective effect of MA-5 was inhibited in PD mice.The effect of MA-5 on PD mice was inhibited using the AMPK inhibitor Compound C intervention.Compared with the MA-5 + MPTP group,PD mice after Compound C pretreatment had decreased motor ability,decreased TH protein expression,decreased expression of mitophagy-related indicators LC3-II/LC3-I,PINK,and Parkin,increased p62 expression,increased levels of proinflammatory cytokines IL-6,IL-1β,and TNF-α in brain tissue,inhibited p-AMPK protein expression,and increased p-m TOR protein expression.【Conclusion】MA-5 can improve motor symptoms and inhibit neuroinflammation in Parkinson’s disease mice.The mechanism may be related to the enhancement of mitophagy by MA-5 activating the AMPK pathway.