Oxidative Stress Attenuation And Apoptosis Suppression By Ultrasound-Assisted Ajuga Bracteosa Extract In Neuronal Cells

Abstract

Ultrasound-assisted extract of Ajuga bracteosa was evaluated for neuroprotective efficacy in mechanistic in vitro models of oxidative stress. SH-SY5Y neuronal cells were exposed to hydrogen peroxide (H₂O₂) after pre-treatment with the extract (25–200 µg/mL). Cell viability (MTT), intracellular reactive oxygen species (DCFH-DA), lipid peroxidation (MDA/TBARS), antioxidant enzymes (SOD, catalase), apoptosis (Annexin V/PI), and key signaling proteins (p-Akt/Akt, Bcl-2, Bax, p53) were quantified. The extract dose-dependently preserved viability and suppressed ROS and MDA formation, while restoring SOD and catalase activities. Flow cytometry showed reduced early and late apoptosis with increased live cell fractions. Western blotting revealed reversal of H₂O₂-induced signaling perturbations through elevation of p-Akt/Akt and Bcl-2 and down-regulation of Bax and p53. These effects approached the antioxidant comparator N-acetylcysteine at higher extract concentrations. The findings suggest that A. bracteosa confers neuroprotection by dual mechanisms: direct redox modulation and reinforcement of endogenous defenses, together with anti-apoptotic signaling via PI3K/Akt. Given the centrality of oxidative stress and apoptosis in neurodegenerative disorders, ultrasound-assisted A. bracteosa extract warrants further investigation, including bioactive characterization, pharmacokinetics, and in vivo validation. This study provides mechanistic evidence supporting the development of A. bracteosa as a candidate phytopharmaceutical for neuroprotection.