卵清转铁蛋白（OVT）已经被确认具有抗炎活性，然而其在胃黏膜炎症中的效应和机制仍不清楚。本研究旨在调查OVT对肿瘤坏死因子-α（TNF-α）诱导的胃上皮细胞（GES-1）炎症反应的效应和机制。采用酶联免疫吸附法（ELISA）确定炎症细胞因子的水平，并通过RNA测序探索其抗炎作用的潜在途径，然后通过西方印迹和途径抑制剂验证。结果显示，OVT在50-400 μg/mL的浓度下对GES-1细胞没有毒性。此外，100 μg/mL的OVT明显降低了白细胞介素（IL）-8、IL-6和TNF-α的分泌，分别降低了63.02%（630.09/1703.98）、35.53%（935.81/1451.43）和36.19%（964.60/1511.63）。RNA测序结果显示，OVT显著影响了丝裂原活化蛋白激酶（MAPK）和核因子κB（NF-κB）途径的激活，这一结果得到了p-IKK、p-IκB、p-P65、p-ERK、p-JNK和p-P38蛋白的水平验证。经过NF-κB和MAPK途径抑制剂孵育后，GES-1细胞释放的IL-8含量进一步证实OVT抑制了这两个途径的激活。总之，这些结果表明OVT是一种具有治疗胃炎潜力的天然蛋白质。

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.