Journal of Traditional Chinese Medical Sciences (Jan 2022)
Mechanism of mitochondrial protection by the Buyin Qianzheng formula in a Parkin overexpression cell model
Abstract
Objective: To identify the molecular mechanisms of the effects of the Buyin Qianzheng formula (BYQZF) on the mitochondrial dynamics in a Parkin overexpression Parkinson's disease (PD) cell model. Methods: First, a stable Parkin overexpression cell model was constructed using plasmid transfection. Then, we examined the protective effect of BYQZF on the mitochondrial dysfunction of the Parkin overexpression PD cell model induced by neurotoxin 1-methyl-4-phenylpyridinium ion (MPP+). The mRNA expression level of Parkin was evaluated using real-time quantitative PCR. The cell survival rate was detected using the Cell Counting Kit-8 assay. We evaluated the cellular adenosine triphosphate (ATP) levels using luciferase assays. A laser scanning confocal microscope was used to observe the mitochondrial morphology, activity, and mitochondrial membrane potential (ΔΨm). Western blot was conducted to evaluate the levels of the fusion proteins mitofusin1, mitofusin2, optic atrophy 1, dynamin-related protein 1, and mitochondrial fission protein 1. Results: Parkin overexpression attenuated MPP+ -induced mitochondrial damage, increased mitochondrial activity and ΔΨm. BYQZF increased the survival of MPP+-induced cells that overexpressed Parkin and upregulated the mitochondrial form factor and activity. It also inhibited a decrease in the ΔΨm and ATP levels. These findings suggested that BYQZF protected against MPP+-induced mitochondrial dysfunction and enhanced the protective effect of Parkin overexpression. Furthermore, the formula upregulated the expression of the fusion proteins mitofusin1, mitofusin2, and optic atrophy 1 (closely related to mitochondrial quality remodeling), and reduced the expression of the fission protein dynamic-related protein 1, as well as mitochondrial fission protein 1. Conclusion: The mechanism by which BYQZF increased the mitochondrial protective effect of Parkin gene overexpression in MPP+ -induced cells may be related to improving mitochondrial function and regulating the balance of mitochondrial division and fusion proteins.
