Abstract
Bisphenol A (BPA), an environmental pollutant, can cause multiple organ tissue damage by inducing oxidative stress. Cineole (CIN) is a terpene oxide existing in a variety of plant essential oils, which has anti-inflammatory, analgesic, and antioxidant effects. This study examined the effects of 200 nM BPA and 20 μM CIN on apoptosis, autophagy, and immunology in grass carp hepatocytes (L8824). The treatments were categorized as NC, CIN, BPA + CIN, and BPA. The findings demonstrated that BPA exposure could increase ROS levels and oxidative stress-related indicators, decrease the expression of the Nrf2/keap1 pathway and the Wnt/β-catenin pathway, increase the expression of genes involved in the apoptotic pathway (Bax and Caspase3), and decrease the expression of the anti-apoptotic gene Bcl-2 by lowering mitochondrial membrane potential. BPA also reduced the expression of genes linked to autophagy (ATG5, Beclin1, LC3). Changes in immunological function after BPA exposure were also shown by changes in the amounts of antimicrobial peptides (HEPC, β-defensin, LEAP2) and cytokines (INF-γ, IL-1β, IL-2, and TNF-α). After the co-treatment of CIN and BPA, CIN can inhibit BPA-induced apoptosis and recover from autophagy and immune function to a certain extent by binding to keap1 to exert an anti-oxidative regulatory effect of Nrf2 incorporation into the nucleus. Molecular docking provides strong evidence for the interaction of CIN ligands with keap1 receptors. Therefore, these results indicated that CIN could inhibit BPA-induced apoptosis, autophagy inhibition and immunosuppression in grass carp hepatocytes by regulating the Wnt/β-catenin pathway with Nrf2/keap1/ROS. This study provided further information to the risk assessment of the neuroendocrine disruptor BPA on aquatic organisms and offered suggestions and resources for further research into the function of natural extracts in the body's detoxification process.
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