Study of the mechanism underlying the role of PINK1/Parkin in the formic acid‐induced autophagy of PC12 cells

Abstract

This study aimed to explore PINK1/Parkin's role in methanol metabolite formic acid‐induced autophagy in PC12 cells and provide a theoretical basis for elucidating methanol‐induced neurotoxicity. After treatment with different formic acid concentrations, we observed the morphology and mitochondria of PC12 cells. We used an ultra‐micro enzyme kit to detect the mitochondrial Na+‐K+‐ATPase and Ca2+‐Mg2+‐ATPase activities; a JC‐1 kit to detect changes in the mitochondrial membrane potential (MMP); MDC staining to detect the autophagy levels; and western blotting to measure the expression levels of the mitochondrial marker protein COX IV and the autophagy‐related proteins Beclin1, P62 and LC3II/LC3I, and the mitochondrial and cytoplasmic levels of PINK1, Parkin and P‐Parkin. Compared with the control group, the mitochondrial diameters, the mitochondrial Na+‐K+‐ATP and Ca2+‐Mg2+‐ATPase activities, the MMP, and the COX IV expression levels decreased significantly (P < 0.05). The fluorescence signal intensity (indicating autophagy); relative Beclin1 and LC3II/LC3I protein expression levels; and relative mitochondrial PINK1, Parkin and P‐Parkin levels increased significantly, and the relative P62 protein expression levels and relative cytoplasmic PINK1, Parkin and P‐Parkin levels decreased significantly (P < 0.05) compared with the control group. Thus, formic acid alters mitochondrial morphology, causes mitochondrial dysfunction, affects the PINK/Parkin pathway and, thus, activates the process of mitochondrial autophagy.