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

Abstract

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-1 cells. Additionally, 100 μg/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-κB) pathways, which was verified by the levels of p-IKK, p-IκB, p-P65, p-ERK, p-JNK, and p-P38 protein. IL-8 contents released by GES-1 cells after incubation with inhibitors of NF-κB 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.

Pongamol Alleviates Neuroinflammation and Promotes Autophagy in Alzheimer’s Disease by Regulating the Akt/mTOR Signaling Pathway

“…As a key activator of inflammatory processes, NF-κB regulates the expression of various inflammatory mediators as well as cytokines in a variety of cell types. 36 Activated NF-κB is transferred to the nucleus and forms a complex with ribosomal protein S3 (RPS3), which then binds to DNA binding sites and regulates the transcription of target genes. 37 In order to explore the antiinflammatory mechanism of pongamol, we detected the expression of related proteins in the NF-κB signaling pathway.…”

“…In this research, we found that pongamol decreased the expression of inflammatory factors. As a key activator of inflammatory processes, NF-κB regulates the expression of various inflammatory mediators as well as cytokines in a variety of cell types . Activated NF-κB is transferred to the nucleus and forms a complex with ribosomal protein S3 (RPS3), which then binds to DNA binding sites and regulates the transcription of target genes .…”

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“…Other than promoting calcium absorption by the body, OVT also stimulated the proliferation and differentiation of osteoblasts and inhibition of osteoclast genesis in vitro, demonstrating that OVT was capable of preventing osteoporosis by mediating bone formation [60] . OVT promoted the maturation of intestinal dendritic cells in mice and exerted immunomodulatory effects against intestinal immunosuppressed mice by regulating the balance of Th1 and Th2 through the mitogen-activated protein kinase (MAPK) signaling pathway [61] . A recent study has investigated the effects of OVT on gastritis and the regulatory mechanisms on gastric epithelial cells (GES-1).…”