Neuroprotective Effect of Magnoflorine Through Inhibition of Caspase-3-Mediated Neuronal Apoptosis: In Silico to In Vitro Validation.
Abstract
Neurodegeneration is a progressive biological phenomenon that is significantly driven by neuronal apoptosis. The current study aimed to investigate the neuroprotective potential of Magnoflorine, a natural alkaloid, by inhibiting Caspase-3-mediated neuronal apoptosis induced by Kainic acid. The caspase-3-inhibiting property of Magnoflorine was initially predicted using in silico methods such as molecular docking, MMGBSA analysis, and molecular dynamics (MD) simulations, taking Kainic acid as the reference neurotoxic ligand. Molecular docking and MMGBSA analysis showed that Magnoflorine has a higher binding affinity for caspase-3 than that of Kainic acid, which predicts the potential of Magnoflorine to inhibit caspase-3 expression. MD simulations further highlighted this potential through the greater stability of the Caspase-3-Magnoflorine complex than the Caspase-3-Kainic acid complex over 200 ns of simulations. It was finally evaluated through multiple in vitro techniques such as the MTT cell viability assay, LDH cytotoxicity assay, Annexin V-PI apoptosis assay, and immunocytochemistry performed on the retinoic acid-differentiated SH-SY5Y cells. MTT and LDH assays revealed that Magnoflorine significantly improved the viability of differentiated SH-SY5Y cells by reducing Kainic acid-induced cell death and cytotoxicity. The apoptosis assay also showed that Magnoflorine significantly reduced the Kainic acid-induced apoptosis in differentiated SH-SY5Y cells. Inhibition of Caspase-3 by Magnoflorine became evident during immunocytochemistry through the reversal of the Kainic acid-induced increase in Caspase-3 expression in differentiated SH-SY5Y cells, as observed from immunofluorescence intensity. The present study suggests that Magnoflorine is emerging as a valuable therapeutic agent for neurodegeneration by alleviating neuronal apoptosis, which requires further validation through in vivo studies.
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