Title:PIASA, A Novel Peptide, Prevents SH-SY5Y Neuroblastoma Cells against
Rotenone-induced Toxicity
Volume: 16
Author(s): Ahmed Sha Sulthana, Rengasamy Balakrishnan, Mani Renuka, Thangavel Mohankumar, Dharmar Manimaran, Kuppamuthu Arulkumar and Elangovan Namasivayam*
Affiliation:
- Department of Biotechnology, School of Bioscience, Periyar University, Salem, Tamilnadu- 636011, India
Keywords:
Parkinson’s disease, SH-SY5Y cells, peptide, cell viability, apoptosis, degeneration.
Abstract:
Background and Objective: This investigation explores 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 were progressed against mitochondrial
respiratory complex I (MCI). The in-silico analysis showed PIASA to have interaction with 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),
the expressions of BAX, Bcl-2, cyt c, pro-caspase 3, and caspase 3, -6, -8, -9, and cleaved caspase
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 expression, while downregulating the Bcl-2 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 the future.
Conclusion: Collectively, our results propose that PIASA mitigated rotenone-stimulated oxidative
stress, mitochondrial dysfunction, and apoptosis in rotenone-induced SH-SY5Y cells.
