Silibinin attenuates 3-nitropropionic acid-induced ovarian toxicity by alleviating oxidative stress and granulosa cell apoptosis.

3-Nitropropionic acid (3-NP), a mycotoxin present in various plants and fungi, poses significant reproductive health risks to animals and humans through food chains contamination. This study aimed to explore the protective effects and mechanisms of silibinin, a bioactive flavonoid derived from the herbal plant Silybum marianum, against 3-NP-induced reproductive toxicity. Our findings demonstrated that silibinin treatment significantly alleviated 3-NP-induced ovarian follicular atresia and preserved ovarian histoarchitecture. Furthermore, it attenuated oxidative stress induced by 3-NP. Molecular analyses through qPCR and Western blot revealed that silibinin upregulated Bax and Caspase-3 expressions while downregulating Bcl-2 expression in the ovaries compared to the 3-NP group. Immunohistochemistry analysis of CASPASE-3 demonstrated that silibinin significantly inhibited 3-NP-induced apoptosis, predominantly in granulosa cells. Additionally, Western blot analyses showed that silibinin suppressed 3-NP-induced activation of JNK and ERK phosphorylation, downregulated KEAP1 expression, and upregulated NRF2 nuclear translocation in the ovary. Overall, our results indicate that silibinin effectively alleviated 3-NP-induced ovarian oxidative damage by modulating the JNK/ERK signaling pathways and activating the KEAP1/NRF2 signaling pathway. These findings suggest that silibinin may have potential therapeutic application for mitigating 3-NP-induced reproductive toxicity in animal husbandry and the veterinary industry.