The Role Of Transient Potential Receptor Channel TRPA1/TRPV1 In Cigarette Induced Airway Epithelial Cell Injury

Objective The purpose of this study was to elucidate the role and mechanism of TRPA1 and TRPV1 in cigarette smoke extract(CSE)induced airway epithelial cell injury model.Methods Bronchial epithelial cells(Bease-2b cells)were co-cultured with 10%CSE and pretreated with A967079(100?m,TRPA1 inhibitor),AMG9810(100p?m,TRPVlinhibitor),and A967079(100?m)+AMG9810(100?m).Intracellular Ca2+level,oxidative stress,the mRNA level of antioxidant(HO-1,NQ01,MnSOD,CAT)and inflammatory cytokines(IL-1?,IL-8,IL-18,IL-33),mitochondrial fission protein(DRP1 MFF)and fusion protein(OPA1 MFN2),NLRP3 inflammasome and Caspase-1 protein were examined.Results In Beas-2b cells,10%CSE induced the Ca2+inflow and increased intracellular and mitochondrial oxidative stress,reduced antioxidants mRNA expression and increased inflammatory cytokines mRNA expression.10%CSE increased the protein expression of MFF and DRP1,decreased the protein expression of OPA1,and up-regulated NLRP3 inflammasome and Caspase-1.Pretreatment with A967079 or AMG9810 or combined use of A967079 and AMG9810 can block Ca2+influx,reduce oxidative stress,increase antioxidants mRNA expression,reduce nflammatory cytokines mRNA expression,prevent the imbalance of mitochondrial fission/fusion protein,and down-regulate NLRP3 inflammasome and Caspase-1.The effects of combined use were better than those of single use.Conclusion Both TRPA1 and TRPV1 mediate CSE-induced airway epithelial cell injury by regulating oxidative stress,inflammatory response and mitochondrial damage,and co-inhibition of TRPA1/TRPV1 channels may better inhibit CSE induced bronchial epithelial cell injury.