Citropten attenuates H₂O₂-induced neurotoxicity by modulating redox balance, inflammation, and apoptotic pathways in SH-SY5Y cells.

Abstract

Objectives: The proposed study explores the neuroprotective potential of Citropten, a natural coumarin derivative, against H₂O₂-induced oxidative stress in SH-SY5Y human neuroblastoma cells. H₂O₂ treatment induced significant cytotoxicity, oxidative damage, mitochondrial dysfunction, and inflammation.

Methods: SH-SY5Y cells were exposed to H₂O₂ to induce oxidative stress, followed by treatment with Citropten. Cell viability was measured using the MTT assay, and oxidative damage was assessed via LDH release, ROS generation, lipid peroxidation, and glutathione reductase (GR) activity. Mitochondrial membrane potential (MMP) was evaluated by flow cytometry. Inflammatory markers were quantified using ELISA, and apoptosis was determined by acridine orange (AO)/ethidium bromide (EB) staining and flow cytometry.

Key findings: Citropten treatment significantly restored cell viability and reduced intracellular ROS levels by 63%, lipid peroxidation by 36% and LDH release by 44.7%, indicating improved membrane integrity. Citropten also preserved MMP (with a 79% restoration) and elevated GR activity. Inflammatory responses were attenuated with a decrease in NF-κB, IL-1β, IL-6, and TNF-α levels. Apoptotic cell death was markedly diminished, as confirmed by AO/EB staining and flow cytometry.

Conclusions: Citropten demonstrated significant antioxidant, anti-inflammatory, and anti-apoptotic properties, highlighting its potential as a promising neuroprotective agent for mitigating oxidative stress-associated neuronal damage and possibly treating neurodegenerative diseases.