Ovotransferrin Inhibits TNF-α Induced Inflammatory Response in Gastric Epithelial Cells via MAPK and NF-icB Pathway

摘要

Ovotransferrin(OVT)has been confirmed to have anti-inflammatory activity.However,its effect and mechanism on gastric inflammation are unclear.In this study,the effect and mechanism of the OVT on the tumor necrosis factor-α(TNF-α)induced inflammatory response in gastric epithelial cells(GES-1)were investigated.The enzyme linked immunosorbent assay(ELISA)was used to determine the levels of inflammation cytokines,followed by RNA sequencing to explore the potential pathways of its anti-inflammatory effect,and then it was validated by Western blotting and pathways inhibitors.Results showed that the OVT at concentrations of 50-400 μg/mL displayed nontoxicity against GES-l cells.Additionally,100/Wg/mL of OVT obviously reduced the secretion of interleukin(IL)-8,IL-6,and TNF-α by 63.02%(630.09/1703.98),35.53%(935.81/1451.43),and 36.19%(964.60/1511.63),respectively.The results of RNA sequencing exhibited that the OVT significantly influences the activation of mitogen-activated protein kinase(MAPK)and the nuclear factor kappa-light-chain enhancer of activated B cell(NF-kB)pathways,which was verified by the levels of p-IKK,p-IxB,p-P65,p-ERK,p-JNK,and p-P38 protein.IL-8 contents released by GES-l cells after incubation with inhibitors of NF-kB and MAPK pathways further confirmed that OVT hindered activation of these two pathways.Collectively,these results suggested that OVT was a natural protein with the potential to treat gastric inflammation.