Chlorogenic acid alleviates IPEC-J2 pyroptosis induced by deoxynivalenol by inhibiting activation of the NF-κB/NLRP3/caspase-1 pathway

Abstract

Abstract Background Deoxynivalenol (DON) is a mycotoxin that severely pollutes feed ingredients, and methods for reducing DON toxicity have become a significant research direction. Chlorogenic acid (CGA) is an active polyphenol found in some plants, which has anti-inflammatory and antioxidant properties and a protective effect on animal intestinal health. The effects of CGA on DON-induced pyroptosis in the intestinal porcine epithelial cell line-J2 (IPEC-J2) and its potential mechanism were explored in this study. Results IPEC-J2 cells viability and membrane integrity were inversely correlated with DON concentration. Compared to those in the group treated with DON alone at 2,500 ng/mL, pretreatment with 80 μmol/L CGA for 4 h significantly improved cell viability ( P < 0.01), and the alleviation of typical pyroptotic symptoms induced by DON were observed, including reduced cellular DNA fragmentation, decreased release of lactate dehydrogenase (LDH), normalized ROS levels, restoration of extracellular Ca 2+ and K + contents to normal levels ( P < 0.01 ), as well as suppressed the enzyme activities of caspase-1 and caspase-4 ( P < 0.01). Additionally, the mRNA expression levels of TNF , MDP , NOD2 , TLR4 , ASC and GSDMD were significantly improved ( P < 0.01), while both mRNA and protein expression levels of NF-κB, NLRP3, caspase-1, IL-1β and IL-18 were significantly upregulated ( P < 0.01) in the CGA + DON group, compare to those in the DON group. Conclusion Pretreatment with 80 μmol/L CGA for 4 h effectively alleviated pyroptosis in IPEC-J2 cells induced by 2,500 ng/mL of DON through inhibiting activation of the NF-κB/ NLRP3/capase-1 pathway.