Hesperidin ameliorates hepatocyte cell cycle disruption, apoptosis, and necroptosis in mice with nonalcoholic fatty liver disease via the VEGFA-PI3K/AKT pathway

Background and purpose: The annual rise in the worldwide incidence of nonalcoholic fatty liver disease (NAFLD) continues; however, there remains a lack of safe and well-proven specific pharmacological treatments for this condition. Hesperidin (HDN), a citrus-derived flavonoid, displays antioxidant properties, and research indicates its capacity to mitigate NAFLD.

Methods: This study developed both in vivo and in vitro high-fat models using high-fat diet (HFD)-induced mice and palmitoleic acid (PO)-stimulated Alpha Mouse Liver 12 (AML12) cells, respectively. We investigated the protective mechanisms of HDN against HFD-induced NAFLD through a comprehensive set of experimental approaches, including pathological examination, RNA Sequencing, Western Blot, immunofluorescence (IF), flow cytometry, acridine orange/ethidium bromide (AO/EB) dyeing, molecular docking, co-immunoprecipitation (Co-IP), and Oil Red O staining. Additionally, siRNA-mediated gene silencing and pharmacological treatment with 740 YP were used to validate the in vitro findings.

Results: Biochemical analysis demonstrated that HDN intervention effectively reduced lipid accumulation in NAFLD. Bioinformatics analysis identified vascular endothelial growth factor A (VEGFA) as a key mediator through which HDN exerts its protective effects against NAFLD. Western Blot and immunofluorescence (IF) analyses revealed that HDN significantly alleviated cell cycle dysregulation, Apoptosis, and Necroptosis in NAFLD mice. Molecular docking and Co-IP results showed that HDN activates the VEGFA-PI3K/Akt signaling pathway to ameliorate NAFLD.

Conclusion: HDN alleviates NAFLD by restoring VEGFA-PI3K/Akt signaling, thereby stabilizing the cell cycle and suppressing Apoptosis and Necroptosis.

Apoptosis; Hesperidin; NAFLD; PI3K/AKT; VEGFA.