Piperonal protects neuron-like and retinalpigment epithelial (RPE) cells from oxidative stress and apoptosis through inhibition of α-synuclein aggregation

Protein fibrillation is a crucial process in the onset of several neurodegenerative and retinal disorders due to the formation of cytotoxic species. Because of their capacity to prevent protein aggregation, small molecules have the potential to be appointed as therapeutic agents. Here, we examined the inhibitory impacts of the piperonal as a carbaldehyde-derived compound on the fibrillation process of α-synuclein and underlying cytotoxicity against neuron-like (PC12) and human retinal pigment epithelial (RPE) (ARPE-19) cells. The results showed that the values of k app and lag time of α-synuclein were modulated with piperonal. Moreover, ANS fluorescence intensity analysis indicated that piperonal can inhibit the formation of a molten global (misfolded) state of α-synuclein, which is a necessary step in the formation of protein amyloid fibrils. Congo red absorption and circular dichroism spectroscopy also verified the inhibition of β-sheet structure formation after treatment of α-synuclein with piperonal. Furthermore, theoretical studies displayed that piperonal interacts with VAL40:HN, GLU35:O, VAL40, and LYS43 amino acid residues and forms a complex. In addition, cytotoxicity assays demonstrated that piperonal as a safe small molecule could mitigate the induced cytotoxicity by α-synuclein amyloids in PC12 and ARPE-19 cells through reduction of ROS and Bax/Bcl2 mRNA overexpression. Taken together, these outcomes showed that piperonal as a natural aldehyde compound can inhibit α-synuclein fibrillation and underlying cytotoxicity which may be developed for potential therapeutic applications in vivo.