目的 探讨佛手柑内酯干预溃疡性结肠炎的潜在分子机制，分析其与m⁶A RNA甲基化相关因子表达变化之间的关联。方法 整合网络药理学和疾病数据库筛选佛手柑内酯抗溃疡性结肠炎的候选靶点，构建蛋白相互作用（PPI）网络并进行富集分析；基于GEO队列（GSE206285）分析m⁶A调节因子在溃疡性结肠炎组织中的差异表达模式。采用分子对接技术评估佛手柑内酯与m⁶A核心蛋白的结合潜能。体外利用葡聚糖硫酸钠诱导NCM460肠上皮细胞建立损伤模型，观察佛手柑内酯对细胞存活率的影响，并检测WTAP、METTL3、METTL14、FTO、YTHDF1、YTHDF2等关键m⁶A因子mRNA表达变化。结果 网络药理学共筛选获得佛手柑内酯干预溃疡性结肠炎的潜在靶点47个，核心靶点以雌激素受体1（ESR1）和细胞色素P450家族（如CYP3A4）为主；主要富集于细胞色素P450介导的外源物代谢、脂质代谢、花生四烯酸代谢及药物代谢等通路。临床队列（GSE206285）分析显示，溃疡性结肠炎组织中METTL3、WTAP表达显著上调，METTL14、YTHDF2表达显著下调（P＜0.001）。分子对接结果表明，佛手柑内酯对核心m⁶A调控蛋白具备潜在亲和力，其中与METTL3、METTL14结合能最强，与WTAP、FTO、YTHDF1/2等呈中等亲和力。体外实验证实，在给予佛手柑内酯干预后，1.25～40 μmol/L佛手柑内酯可显著提高葡聚糖硫酸钠受损细胞的存活率（P＜0.05、0.001）。佛手柑内酯（1.25、2.5、5 μmol/L）组WTAP、METTL3、YTHDF1表达均显著下调（P＜0.001），佛手柑内酯（2.5、5 μmol/L）组YTHDF2、FTO表达显著升高（P＜0.01、0.001），佛手柑内酯（5 μmol/L）组METTL14表达显著上调（P＜0.001）。结论 佛手柑内酯可能通过调节外源物代谢、脂质代谢及炎症相关通路并纠正m⁶A修饰相关因子失衡来发挥抗溃疡性结肠炎作用。METTL3、METTL14、WTAP等关键m⁶A调控蛋白可能是其发挥表观遗传调控的潜在靶点。

Objective To explore the potential molecular mechanisms by which bergapten intervenes in ulcerative colitis, and to preliminarily analyze its association with expression changes of m⁶A RNA methylation–related regulators. Methods Network pharmacology was integrated with disease databases to screen candidate targets of bergapten against ulcerative colitis, and PPI network was constructed and subjected to enrichment analysis. Differential expression patterns of m⁶A regulators in ulcerative colitis tissues were analyzed based on the GEO cohort GSE206285. Molecular docking was employed to evaluate the binding potential between bergapten and core m⁶A proteins. In vitro, a dextran sodium sulfate-induced injury model was established in NCM460 intestinal epithelial cells to observe the protective effect of bergapten on cell viability, and to examine expression changes of key m⁶A factors, including WTAP, METTL3, METTL14, FTO, YTHDF1, and YTHDF2. Results Network pharmacology identified 47 potential targets of bergapten intervention in ulcerative colitis. The core targets were mainly represented by ESR1, and members of the cytochrome P450 family, such as CYP3A4. These targets were mainly enriched in cytochrome P450-mediated xenobiotic metabolism, lipid metabolism, arachidonic acid metabolism, drug metabolism, and related pathways. Analysis of the clinical cohort GSE206285 showed that METTL3 and WTAP were significantly upregulated in ulcerative colitis tissues, whereas METTL14 and YTHDF2 were significantly downregulated (P < 0.001). Molecular docking results indicated that bergapten showed potential affinity for core m⁶A regulatory proteins, with the strongest binding energies observed for METTL3 and METTL14, while WTAP, FTO, YTHDF1/2 and other proteins showed moderate affinity. In vitro experiments have confirmed that bergapten 1.25—40 μmol/L can significantly increase the survival rate of cells damaged by dextran sulfate sodium (P < 0.05, 0.001). Expressions of WTAP, METTL3, and YTHDF1 in the bergapten (1.25, 2.5, 5 μmol/L) groups were significantly downregulated (P < 0.001), while the expressions of YTHDF2 and FTO in the bergapten (2.5 and 5 μmol/L) groups were significantly increased (P < 0.01, 0.001), and the expression of METTL14 in the bergapten (5 μmol/L) group was significantly upregulated (P < 0.001). Conclusion Bergapten may exert its anti- ulcerative colitis effects by regulating xenobiotic metabolism, lipid metabolism, and inflammation-related pathways, as well as by correcting the imbalance of the m⁶A modification network. Key m⁶A regulatory proteins, such as METTL3, METTL14, and WTAP may serve as potential targets through which bergapten exerts epigenetic regulatory effects.

R285.5;R286.5

国家中医药管理局科技司科技共建项目（GZY-KJS-GD-2025-093）；广东省基础与应用基础研究基金项目（2023A1515011699）；中山市社会公益与基础研究专项（2025B2007）；广州中医药大学医院联合基金项目（GZYZS2024U08）