Hesperidin alleviated dendritic spines through inhibiting ferritinophagy via HERC2-NCOA4 ubiquitination in CUMS mice.

Background: Depression has been linked to ferritinophagy-induced synaptic damage, which affects the emotional circuitry and can ultimately lead to depressive symptoms. It has been suggested that Hesperidin might improve depression disorders. However, the relationship between the therapeutic effects of the sympathetic nervous system in alleviating depression-like behaviors and ferritinophagy is still unknown.

Purpose: The objective of this study is to investigate the possible impact of Hesperidin in alleviating dendritic spines through the inhibition of ferritinophagy via HERC2-NCOA4 ubiquitination in mice exposed to chronic unpredictable mild stress (CUMS).

Methods: C57BL/6 and NCOA4 mice were exposed to CUMS for 42 days. During the last 3 weeks of the CUMS procedure, the mice were administered Hesperidin (50, 100, 200 g/kg/d) or fluoxetine (10 mg/kg/d) once daily. Following the behavioral tests, Golgi staining, tissue iron concentration test, and perls staining were conducted to assess the therapeutic effect of Hesperidin. Additionally, ultrahigh-performance liquid chromatography-high-resolution tandem mass spectrometry (UPLC-Q-TOF/MS) and ultrahigh-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was utilized to analyze the metabolic components of Hesperidin in both blood and brain tissue. To investigate mechanism of Hesperidin, the cells were subjected to different concentrations of Hesperidin (25, 50, 100 µM), its metabolites (Eriodictyol (10, 20, 50 µM), Homoeriodictyol (0.1, 0.5, 1 µM)) and si-HERC2. Furthermore, HERC2-NCOA4 ubiquitination, and ferritinophagy-related proteins was explored through techniques such as western blot, immunofluorescence, co-immunoprecipitation, and molecular docking.

Results: Hesperidin has demonstrated the potential to alleviate symptoms of depression by regulating dendritic spines through the inhibition of NCOA4-ferritinophagy, while NCOA4 overexpression could reverse these results. Importantly, the content of Hesperidin metabolites (Homoeriodictyol and Eriodictyol) was relatively high in brain tissue. The Hesperidin and its metabolites, Eriodictyol and Homoeriodictyol, were able to regulate GluR2 and SYN protein expression. Additionally, they inhibited ferritinophagy involving NCOA4, P62, LC3, and FTH. but this phenomenon was reversed by si-HERC2 following Hesperidin and its metabolite administration. Furthermore, the binding of HERC2 and NCOA4 protein was found to be inhibited by Hesperidin and its metabolites.

Conclusions: Hesperidin alleviated dendritic spines through inhibiting ferritinophagy via HERC2-NCOA4 ubiquitination in CUMS mice.