目的 探究小檗碱促进肛瘘术后愈合的分子机制。方法 通过GEO数据集（GSE157020）收集肛瘘患者差异基因，利用PharmMapper、SwissTargetPrediction、TCMSP数据库获取小檗碱靶点，取两者交集筛选关键靶点；借助DAVID数据库进行基因本体（GO）和京都基因和基因组百科全书（KEGG）富集分析，通过Cytoscape构建“成分–靶点–疾病”网络，利用AutoDockTools进行分子对接，通过iMODS在线服务器开展黄连素与关键靶点的分子动力学模拟，并进行体外实验验证。结果 筛选得到11个关键靶点[过氧化物酶体增殖物激活受体δ（PPARD）、中链酰基辅酶A脱氢酶（ACADM）、核受体亚家族1H组成员2（NR1H2）等]。GO分析显示靶点涉及脂质代谢、炎症反应调节等生物学过程，KEGG分析涉及花生四烯酸代谢、PPAR信号通路等；分子对接显示小檗碱与ACADM、PPARD、NR1H2的结合能分别为-11.0、-9.3、-7.0 kcal/mol；iMODS模拟表明复合物构象稳定性较高。通过体外实验验证小檗碱可逆转肿瘤坏死因子-α（TNF-α）诱导PPARD、ACADM、NR1H2的蛋白和mRNA表达（P＜0.05、0.01）。结论 小檗碱可能通过调控上述关键靶点及其PPAR通路，进而发挥促进肛瘘术后愈合的作用，为临床治疗提供了潜在靶点。

Objective To explore the molecular mechanism by which berberine promotes postoperative wound healing of anal fistula. Methods Differential genes of patients with anal fistula were collected from the GEO dataset (GSE157020). The targets of berberine were obtained using PharmMapper, SwissTargetPrediction, and TCMSP databases, and the intersection of the two was taken to screen key targets. GO and KEGG enrichment analyses were performed with the help of the DAVID database. The “component-target-disease” relationship network was constructed using Cytoscape. Molecular docking was carried out using AutoDockTools, and molecular dynamics simulation of berberine and key targets was conducted through the iMODS online server, and conduct in vitro experiments for verification. Results A total of 11 key targets were screened out (including ACADM, PPARD, NR1H2, etc). GO analysis showed that the targets were involved in biological processes such as lipid metabolism and regulation of inflammatory response. KEGG analysis involved arachidonic acid metabolism, PPAR signaling pathway, etc. Molecular docking showed that the binding energies of berberine with ACADM, PPARD, and NR1H2 were -11.0, -9.3, and -7.0 kcal/mol, respectively. iMODS simulation indicated that the conformation of the complex had high stability. In vitro experiments confirmed that berberine can reverse the TNF-α-induced mRNA and protein expression of PPARD, ACADM, and NR1H2 (P < 0.05, 0.01). Conclusion Berberine may exert a role in promoting post-operative healing of anal fistula by regulating the aforementioned key targets and their associated PPAR pathway, which provides potential targets for clinical treatment.

R285.5

湖北省自然科学基金创新发展联合基金项目（2025AFD310）