Modulation of autophagy and apoptosis processes in experimental models of neuroinflammation

Neuroinflammation represents a key pathological mechanism underlying the onset and progression of several neurodegenerative diseases, including Alzheimer's disease (AD). Persistent activation of innate immune pathways, together with oxidative stress and impaired cell death processes, contributes to neuronal dysfunction and degeneration. In this context, the interplay between inflammasome activation, autophagy, and apoptosis has emerged as a critical determinant of neuronal survival and inflammatory balance in the central nervous system. The present study aimed to investigate the molecular mechanisms involved in neuroinflammatory responses and neurodegeneration, focusing on the modulation of autophagy and apoptosis pathways and the regulatory role of non-coding RNAs. Additionally, the potential neuroprotective effects of two phytocannabinoids, cannabidiol (CBD) and cannabigerol (CBG), were evaluated in experimental models of Alzheimer’s disease–related neurotoxicity. The proposed in vitro model of Alzheimer’s disease (AD) was established by exposing human neuroblastoma SH-SY5Y cells and human microglial HMC3 cells to oligomeric β-amyloid₁₋₄₂. Cellular responses were evaluated using complementary approaches, including MTT assays for cell viability, quantitative RT-PCR for inflammatory and apoptotic gene expression, fluorescence-based detection of reactive oxygen species (ROS), and Western blot analysis of proteins involved in inflammasome activation and autophagy (NLRP3, LC3B, and p62). Drebrin expression, a cytoskeletal protein associated with synaptic plasticity, was further assessed by immunofluorescence. Small RNA-seq combined with bioinformatic analysis was employed to characterise the miRNA expression profile associated with neuroinflammatory conditions. Differential expression analysis and functional enrichment of predicted miRNA target genes allowed the identification of regulatory networks linked to inflammation, oxidative stress, apoptosis, and mitochondrial dysfunction. The results suggest that exposure to β-amyloid induces a marked inflammatory and oxidative response in both neuronal and microglial cells, accompanied by alterations in autophagy-related pathways and activation of apoptotic mechanisms. Treatment with CBD and CBG demonstrated potential neuroprotective effects, reducing oxidative stress and modulating key molecular pathways associated with neuroinflammation and cell death. Moreover, miRNA profiling revealed specific regulatory signatures potentially involved in the modulation of inflammatory and degenerative processes. These findings contribute to a deeper understanding of the molecular mechanisms linking neuroinflammation, autophagy, and apoptosis in neurodegenerative contexts and highlight the potential role of cannabinoids as modulators of neuroinflammatory pathways. Such insights may support the identification of novel therapeutic targets and strategies aimed at slowing or preventing the progression of Alzheimer’s disease.