Zhang Hui, Xu Meili
Department of Gerontology, The Second Xiangya Hospital, Central South University, Changsha, Hunan, P.R. China.
Autoimmunity. 2025 Dec;58(1):2491717. doi: 10.1080/08916934.2025.2491717. Epub 2025 May 19.
The prevalence of inflammatory bowel disease (IBD) has increased recently and lacks curative treatments. The involvement of the N6-methyladenosine (mA) reader YTH N6-methyladenosine RNA binding protein 1 (YTHDF1) in ulcerative colitis (UC)-like model was studied in this study. DSS was employed to induce the UC-like condition both and . TNF-α, IL-1β, IL-6, and IL-8 secretion levels were analyzed by ELISA assay. Cell vitality was determined by CCK8 assay. FOXO1 mRNA mA level was examined using methylated RNA binding protein immunoprecipitation (Me-RIP) assay. The interactions between YTHDF1 and FOXO1 were analyzed by RIP assay. ChIP and dual luciferase reporter assays were used to explore the relationship between FOXO1 and FBW7. YTHDF1, FOXO1, and FBW7 were overexpressed in DSS-induced colon epithelial cells. YTHDF1 downregulation alleviated DSS-induced inflammation and NF-κB signal activation in colon epithelial cells. Mechanically, YTHDF1 increased FOXO1 mRNA stability in an mA manner. YTHDF1 overexpression prevented the inhibition of FOXO1 knockdown on DSS-induced inflammation in colon epithelial cells. In addition, FOXO1 transcriptionally activated FBW7. Moreover, FOXO1 upregulation abolished the inhibitory effect of FBW7 knockdown on DSS-induced inflammation in colon epithelial cells. Animal experiments also showed that YTHDF1 deletion alleviated inflammatory response in UC-like mice. YTHDF1 promoted inflammatory response in the UC-like model by transcriptionally activating FBW7 through regulating mA-dependent FOXO1.
炎症性肠病（IBD）的患病率最近有所上升，且缺乏治愈性治疗方法。本研究探讨了N6-甲基腺苷（m⁶A）阅读器YTH N6-甲基腺苷RNA结合蛋白1（YTHDF1）在溃疡性结肠炎（UC）样模型中的作用。采用葡聚糖硫酸钠（DSS）诱导UC样状态。通过酶联免疫吸附测定（ELISA）分析肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和白细胞介素-8（IL-8）的分泌水平。通过细胞计数试剂盒-8（CCK8）测定法检测细胞活力。使用甲基化RNA结合蛋白免疫沉淀（Me-RIP）测定法检测叉头框蛋白O1（FOXO1）mRNA的m⁶A水平。通过RNA免疫沉淀（RIP）测定法分析YTHDF1与FOXO1之间的相互作用。采用染色质免疫沉淀（ChIP）和双荧光素酶报告基因测定法探讨FOXO1与F-Box和WD重复结构域蛋白7（FBW7）之间的关系。YTHDF1、FOXO1和FBW7在DSS诱导的结肠上皮细胞中过表达。YTHDF1下调减轻了DSS诱导的结肠上皮细胞炎症和核因子-κB（NF-κB）信号激活。机制上，YTHDF1以m⁶A依赖的方式增加FOXO1 mRNA的稳定性。YTHDF1过表达可防止FOXO1敲低对DSS诱导的结肠上皮细胞炎症的抑制作用。此外，FOXO1可转录激活FBW7。而且，FOXO1上调消除了FBW7敲低对DSS诱导的结肠上皮细胞炎症的抑制作用。动物实验还表明，YTHDF1缺失减轻了UC样小鼠的炎症反应。YTHDF1通过调节m⁶A依赖的FOXO1转录激活FBW7，从而促进UC样模型中的炎症反应。
