Chlorogenic acid attenuates deoxynivalenol-induced apoptosis and pyroptosis in human keratinocytes via activating Nrf2/HO-1 and inhibiting MAPK/NF-κB/NLRP3 pathways

Abstract

Abstract Background and aim Deoxynivalenol (DON) is a common mycotoxic contaminant, frequently found in food and feed, causing a severe threat to human and animal health. Because of the widespread contamination of DON, humans involved in agricultural practices may be directly exposed to DON through skin route. Chlorogenic acid (CGA) is a phenolic acid, which has anti-inflammatory and antioxidant properties. However, it is still unclear whether CGA can protect against DON-induced skin damage. Here, the effect of CGA on mitigating damage to human keratinocytes (HaCaT) triggered by DON, as well as its underlying mechanisms were investigated. Methods HaCaT cells were pre-incubated with CGA for 2 h, and then exposed with 0.25 µg/mL DON for 24 h, and cell viability was examined with CCK-8 assay. ROS (reactive oxygen species) was measured by flow cytometry mitochondrial and reactive oxygen species (mtROS) was measured fluorescence microscope. Superoxide dismutase (SOD), glutathione (GSH) and catalase (CAT) were measured by using corresponding kits.Proteins expression levels were determined by Western blot analysis. Results CGA pretreatment for 2 h significantly increased cell viability and reversed DON-induced oxidative stress by improving antioxidant enzyme activities such as SOD, GSH, CAT, reducing mtROS generation and enhancing mitochondrial function through activating Nrf2/HO-1 pathway. Moreover, CGA significantly increased the Bcl-2 protein expression, and decreased the protein expressions of Bax and cleaved caspased-3, and suppressed the phosphorylated of ERK, JNK, NF-κB. In addition, CGA could also inhibited the pyroptosis-related protein expressions including NLRP3, cleaved caspased-1 and cleaved IL-1β. Conclusion Our results suggest that CGA could attenuate DON-induced oxidative stress, inflammation and apoptosis through activating Nrf2/HO-1 pathway and inhibiting MAPK/NF-κB and pyroptosis signaling pathway.