Work

Melatonin protects chicken follicular granulosa cells against oxidative damage-induced apoptosis via inhibiting FOXO1 signaling pathway

Year: 2025

Type: article

Abstract: In laying hens, the age-related decline in egg production efficiency is largely attributed to ovarian dysfunction and increased follicular atresia, which are aggravated by oxidative damage mediated by reactive oxygen species ( ROS ). Melatonin ( Mel ) demonstrates powerful antioxidant capabilities, which are essential in eliminating free radicals and enhancing antioxidant enzymes, crucial for the development of follicles. Forkhead Box Protein O1 ( FOXO1 ), a key member of the FOXO family, plays a vital role in regulating various physiological processes involved in follicle growth, maturation, and atresia. This study focused on examining the impact of Mel on chicken follicular granulosa cells ( CFGCs ) upon oxidative stress and investigated its specific mechanisms. An oxidative stress model was developed using exposure to hydrogen peroxide ( H₂O₂ ) at a concentration of 50 μmol/L for 6 h. This model was based on dose-response curves analyzing cell viability, ROS fluorescence intensity, contents of malondialdehyde ( MDA ), and antioxidant enzyme activities ( SOD ). Subsequently, we explored the effects of Mel on CFGCs under conditions of oxidative stress, discovering that Mel mitigated the oxidative damage and cellular apoptosis triggered by H 2 O 2 . This protective effect was achieved by significantly reducing ( P < 0.05) ROS production and modulating the expression of key genes and proteins. It led to upregulation of catalase ( CAT ), superoxide dismutase isoforms 1 and 2 ( SOD1, SOD2 ), glutathione peroxidase 3 ( GPX3 ), peroxiredoxin 3 ( PRDX3 ), and B-cell lymphoma 2 ( BCL-2 ), but downregulated BCL-2-interacting mediator of cell death ( BIM ), Caspase-3, and BCL-2-associated X protein ( BAX ) upon H 2 O 2 exposure ( P < 0.05). Additionally, the expression levels of FOXO1, a critical gene and protein in the Mel-treated group, were significantly reduced ( P < 0.05), indicating that Mel may safeguard ovarian cells from oxidative damage by suppressing the FOXO1 signaling pathway. To further examine this, we developed models using siRNA against FOXO1 ( si-FOXO1 ) and overexpressing FOXO1 ( pcDNA3.1-FOXO1 ). The results indicated that exposure to Mel significantly increased ( P < 0.05) BCL-2 levels while decreasing FOXO1, BIM, Caspase-3, and BAX expressions in both si-FOXO1 and pcDNA3.1-FOXO1 models. Ultimately, these findings suggest that Mel protects CFGCs against oxidative damage-induced apoptosis through inhibiting FOXO1 signaling pathway.

Source: Poultry Science

DOI: 10.1016/j.psj.2025.105472