Effects of Wenfei Jiangzhuo Formula on PINK1/PARKIN-Mediated Mitochondrial Autophagy via Aβ25-35-Induced PC12 Cell Injury
Objective:To explore the effect of Wenfei jiangzhuo formula on mitochondrial autophagy through Aβ25-35 induced PC 12 cell injury.Methods:PC12 cells were differentiated by recombinant ratβ-NGF,Aβ25-35 intervened to become the AD cell model,the opti-mal concentration and time point of Aβ25-35 model was determined.The low(2％),medium(4％)and high(8％)concentration of Wen-fei jiangzhuo formula drug-containing serum groups were set up.MDC staining and JC-1 staining methods were used to observe the auto-phagosome and mitochondrial membrane potential.Immunofluorescence chemical staining,qPCR and Western Blot were used to detect the mRNA and/or protein expressions of PINK1,PARKIN,Ubl,Ub,p-Ub and LC3B in the cells of each group.Results:The PC 12 cells differentiated byβ-NGF became the morphological structure of hippocampal neurons after six days,and the optimal molding condition of AD cell was 5 μmol/L Aβ25-35 intervention for 48 hours.Compared with the normal control group,the model group had a significant increase in autophagosomes,a significant reduction in mitochondrial membrane potential,and significant increases in mRNA and/or protein expressions of PINK 1,PARKIN,Ubl,Ub,p-Ub,LC3B(P＜0.05).Compared with the model group,the Wenfei jiangzhuo formula drug-con-taining serum had a significant reduction in autophagosomes of model cell,a significant increase in mitochondrial membrane potential,and significant reductions in mRNA and/or protein expressions of PINK1,PARKIN,Ubl,Ub,p-Ub,LC3B(P＜0.05).Conclusion:Wenfei jian-gzhuo formula can attenuate Aβ25-35-induced PC12 cell injury possibly through PINK1/PARKIN mediated mitophagy pathway.
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