Lycium barbarum Cyclic Peptide Regulates NLRP3 and NF-κB Signaling Pathways to Reduce BaP-Induced Inflammatory Airway Epithelial Cell Injury
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Abstract:
In this study, Lycium barbarum L. bioactive peptide (LBP) was isolated by the activity tracking method, and its activity against benzopyrene (BaP)-induced airway epithelial cell injury and the potential underlying mechanism were investigated. The cytotoxicity of LBPs was examined by the CCK-8 method; The inhibitory effects of LBP-1 against BaP-induced secretion of inflammatory factors (TNF-α, IL-1β, IL-6, IL-18, NO and PGE2) in human bronchial epithelial cells (16-HBE cells) were examined by ELISA; Western Blot method was used to examine the effects of LBP-1 and BaP on the expressions of COX-2, iNOS, NLRP3 inflammasomes and NF-κB signaling pathway-related proteins in 16-HBE cells. The results showed that LBP-1 could significantly reduce the decrease of 16-HBE cell viability caused by BaP exposure, and there was insignificant cytotoxicity when the concentration was lower than 1 mmol/L; LBP-1 decreased BaP exposure-induced increase in the secretion of cytokines, and the concentrations of TNF-α, IL-1β, IL-6, IL-18, NO and PGE2 decreased by 34.93%, 27.41%, 31.05%, 35.28%, 51.15% and 27.46% respectively, while the expressions of the key enzymes (COX-2 and iNOS) of PGE2 and NO decreased by 81.72% and 41.70% respectively; Western Blot results showed that LBP-1 could inhibit the phosphorylation of IκBα and p65, reduce the expressions of NLRP3 and apoptosis-associated speck-like protein containing CARD (ASC), thereby inhibiting the activation of NLRP3 and NF-κB signaling pathways. The above results confirmed that LBP-1 could play a role in counteracting BaP-induced inflammatory airway epithelial cell injury through regulating the expressions of inflammatory cytokines and NLRP3 and NF-κB signaling pathway-related proteins.
