PIASA, A Novel Peptide, Prevents SH-SY5Y Neuroblastoma Cells against Rotenone-induced Toxicity

Abstract

Background and Objective: This investigation explored the neuroprotective effect of PIASA a newly designed peptide - VCSVY in in-silico and in opposition to rotenone stimulated oxidative stress, mitochondrial dysfunction, and apoptosis in an SH-SY5Y cellular model. Methods: Docking and visualization of the PIASA and rotenone was progressed against mitochondrial respiratory complex I (MCI). The in-silico analysis showed that PIASA had interaction to the binding sites of rotenone which may reduce the rotenone interaction and its toxicity too. The SH-SY5Y cells were segregated into four experimental groups: Group I: untreated control cells; Group II: rotenone-only (100 nM) treated cells; Group III: PIASA (5 µM) + rotenone (100 nM) treated cells; and Group IV: PIASA-only (5 μM) treated cells. Results: We evaluated the cell viability, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), apoptosis (dual staining technique), nuclear morphological changes (Hoechst staining technique), expression of BAX, Bcl-2, cyt c, pro-caspase 3, and caspases 3, -6, -8, -9 and cleaved caspases 3 by western blot analysis. In SH-SY5Y cells, we further observed the cytotoxicity, oxidative stress and mitochondrial dysfunction in rotenone-only treated cells, whereas pretreatment of PIASA attenuated the rotenone mediated toxicity. Moreover, rotenone toxicity is caused by complex I inhibition, which leads to mitochondrial dysfunction, increased BAX while down regulating the Bcl2 expression and cyt c release and then finally caspases activation. PIASA pretreatment prevented the cytotoxic effects via the normalization of apoptotic marker expressions influenced by rotenone. In addition, pre-clinical studies are acceptable in rodents to make use of PIASA as a revitalizing remedial agent especially for PD in future. Conclusion: Collectively, our results proposed that PIASA mitigated rotenone stimulated oxidative stress, mitochondrial dysfunction, and apoptosis in rotenone induced SH-SY5Y cells.