Hesperetin Reduces Neuronal Death in an Shsy5y Alzheimer’S Model by Inhibiting Inflammation and Apoptosis and Pyroptosis Cell Death Pathways Publisher Pubmed



Shafiee L ; Pishva MS ; Hosseinzadegsn R ; Bahadori Z ; Baziyar P ; Mehboodi M ; Khademee S ; Akbari M ; Motamed M ; Nadimi E
Source: Scientific Reports Published:2025

Abstract

Alzheimer’s disease (AD) features amyloid-β (Aβ)1–42 plaques, neuroinflammation, and neuronal loss. Apoptosis and pyroptosis contribute to AD, with inflammatory cytokines involved. Flavonoids like Hesperetin may reduce Aβ1–42 deposition through anti-inflammatory effects. This study introduces a novel method combining LPS and Aβ1–42 to investigate Hesperetin’s mechanism for potential AD treatments. Using computational and experimental methods, we evaluated the physicochemical properties and their correlation with protein aggregation at the molecular level. Human neuroblastoma SH-SY5Y cells were induced to differentiate and then exposed to Hesperetin (1 µM and 10 µM), LPS (1 µg/mL), and Aβ1–42 (20 µM) for 24 h. The expression levels of pro- (Bak, Bax, and Caspase-3) and anti-apoptotic genes (Bcl-2), pyroptosis-related genes (Caspase-1, Caspase-4, Caspase-5, NLRP3, and GSDMD), and pro-inflammatory cytokines genes (interleukins 6 and 1β, and TNF-α) were analyzed via qRT-PCR. The obtained simulation of our result clearly showed that Hesperetin led to the disintegration of the cross-linked structure of organized Aβ1–42 fibrils. Increased RMSD, Rg, and SASA values ​​might lead to destabilization of Aβ1–42 fibrils in the presence of Hesperetin. Our experimental study also demonstrated that Hesperetin increased cell viability in SH-SY5Y cells induced by LPS and Aβ1–42. Hesperetin effectively reverses the enhanced apoptosis caused by LPS and Aβ1–42. Our findings indicated that Hesperetin significantly reduced the elevated expression levels of pro-inflammatory cytokines in the SH-SY5Y cells induced by LPS and Aβ1–42. Treatment with Hesperetin led to a notable downregulation of the enhanced expression of pyroptotic-related genes in LPS and Aβ1–42 induced cells. The details of the molecular level along with the investigation of the physicochemical properties of Hesperetin regarding the mechanism of destabilization of Aβ1–42 fibrils introduce it as a promising therapeutic agent for AD management. Our experimental findings also indicate that Hesperetin is a compound that prevents neuronal death by reducing inflammation and inhibiting apoptosis and pyroptosis. © The Author(s) 2025.