Formulation and Evaluation of Astaxanthin-Loaded Invasomes as Therapeutic Approaches for Alzheimer's Disease Induced in Rats: Role of SIRT-1/BDNF/miRNA-134/GSK-3β Signaling.

Alzheimer's disease (AD) is a progressive age-dependent neurodegenerative disorder associated with oxidative brain damage, disrupted neuronal transmission, memory loss, and behavioral changes, with aluminum being a key environmental risk factor that exacerbates its effect. The aim of this study is to enhance the therapeutic potential of astaxanthin (AST) in Alzheimer's disease by formulating it into invasomal carriers, with special emphasis on SIRT-1/BDNF/miRNA-134/GSK-3β signaling in an AD-like rat model caused by aluminum chloride (AlCl) at a dose of 100 mg/kg/day for 60 days. Optimum AST-loaded invasomes (AST-LI) were prepared using a formulation of phospholipid: ethanol: cineole as 300 mg: 0.3 ml: 0.1 ml for the production of stable vesicles with high entrapment efficiency and negative zeta potential indicating good stability and de-aggregation. As a SIRT-1 activator, AST-LI supplementation improved learning and memory by alleviating the brain redox status (reduced glutathione; GSH, malondialdehyde; MDA), mitochondrial dysfunction, and inflammatory response linked to amyloid β (Aβ) clearance and GSK-3β-mediated p-tau inhibition. It enhanced both spatial and non-spatial short-term memory in rats and restored neurotransmitter levels by raising serotonin and reducing acetylcholinesterase (AChE) and monoamine oxidase (MAO) activities in the brain. Furthermore, AST-LI significantly restored the brain recovery proteins such as Chemokine C-X3-C motif ligand 1 (CX3CL1), glial fibrillary acidic protein (GFAP), brain-derived neurotrophic factor (BDNF), and miRNA-134. These modulations may underlie the observed improvements in oxidative stress, inflammation, apoptosis, and histological outcomes in the AD-like model. In conclusion, improved AST-LI formulations represent promising therapeutic approaches for AD by modulating SIRT-1/BDNF/miRNA-134/GSK-3β signaling.