目的 通过网络药理学及实验验证探讨苦参碱治疗急性胰腺炎炎症反应的分子作用机制。方法 通过DisGeNET、GeneCards、OMIM数据库获取急性胰腺炎相关靶点，TCMSP数据库筛选苦参碱作用靶点，取交集得到苦参碱治疗急性胰腺炎的潜在靶点。利用DAVID和京都基因与基因百科全书（KEGG）数据库进行基因本体（GO）功能和KEGG通路富集分析，通过String数据库构建蛋白质相互作用（PPI）网络，结合cytoHubba算法筛选核心靶点。采用在线分子对接工具验证苦参碱与核心靶点的结合能力。通过雨蛙肽诱导MPC83细胞损伤模型，CCK-8检测细胞活力，RT-PCR检测炎症因子[白细胞介素（IL-6）、肿瘤坏死因子-α（TNF-α）、转化生长因子-β（TGF-β1）、IL-1β]mRNA表达；通过雨蛙素、脂肪酶联合诱导C57BL/6小鼠急性胰腺炎模型，苏木精–伊红（HE）染色观察胰腺组织病理变化，ELISA检测小鼠血清淀粉酶（AMY）、IL-6、TNF-α水平。结果 网络药理学分析发现苦参碱治疗急性胰腺炎的潜在靶点共30个，富集分析显示，苦参碱治疗急性胰腺炎的靶点显著富集于细胞因子受体结合、炎症反应等GO功能，癌症通路、IL-17信号通路等KEGG通路。PPI网络筛选出前5位核心靶点为IL-6、TNF、TGF-β1、IL-1β、NFκBIA。分子对接显示苦参碱与IL-6、TNF、TGFβ1、IL-1β结合效能稳定。细胞实验表明，苦参碱可显著提高损伤细胞活力（P＜0.05），下调IL-6、TNF-α、TGF-β1、IL-1β mRNA表达（P＜0.05）。动物实验显示，苦参碱高剂量组显著降低急性胰腺炎小鼠血清AMY、IL-6、TNF-α水平（P＜0.05、0.01），减轻胰腺组织水肿、坏死及炎细胞浸润。结论 苦参碱通过靶向IL-6、TNF、TGF-β1、IL-1β等炎症相关基因，抑制炎症反应，改善急性胰腺炎病理损伤，其作用机制与调控细胞因子信号通路密切相关，为急性胰腺炎治疗提供了潜在药物靶点和理论依据。

Objective To explore the molecular mechanism of matrine in alleviating inflammatory responses in acute pancreatitis by network pharmacology and experimental validation. Methods Acute pancreatitis -related targets were obtained from DisGeNET, GeneCards, and OMIM databases. Matrine’s action targets were screened using the TCMSP database, and the intersection of these targets was identified as potential targets for matrine in treating acute pancreatitis. GO functional and KEGG pathway enrichment analyses were performed using DAVID and KEGG databases. A protein-protein interaction (PPI) network was constructed via the String database, and core targets were screened using the cytoHubba algorithm. Online molecular docking tools were used to validate the binding ability of matrine to core targets. A cerulein-induced MPC83 cell injury model and cell viability were detected by CCK-8 assay. The expression of inflammatory cytokines (IL-6, TNF-α, TGF-β1, IL-1β) mRNA was detected by RT-PCR. A cerulein/lipase-induced C57BL/6 mice acute pancreatitis model was used. Pathological changes in pancreatic tissues were observed via HE staining. Serum levels of AMY, IL-6, and TNF-α in mice were measured by ELISA. Results A total of 30 potential targets for matrine in acute pancreatitis treatment were identified. Enrichment analysis showed that these targets were significantly enriched in GO such as cytokine receptor binding, inflammatory response, as well as KEGG pathways such as pathways in cancer, IL-17 signaling pathway. The top 5 core targets screened from the PPI network were IL-6, TNF, TGFβ1, IL-1β, and NFκBIA. Molecular docking showed that matrine had stable binding efficacy with IL-6, TNF, TGFβ1 and IL-1β. Cell experiments showed that matrine significantly enhanced the viability of injured cells (P< 0.05), and downregulated the mRNA expression of IL-6, TNF-α, TGF-β1, and IL-1β (P< 0.05). Animal experiments showed that matrine group significantly reduced serum AMY, IL-6, and TNF-α levels (P< 0.05, 0.01) in acute pancreatitis mice, and alleviated pancreatic tissue edema, necrosis, and inflammatory cell infiltration. Conclusion Matrine inhibits the inflammatory response and improves acute pancreatitis pathological damage by targeting inflammation-related genes such as IL-6, TNF, TGF-β1, and IL-1β. Its mechanism is closely related to regulating cytokine signaling pathways, providing potential drug targets and a theoretical basis for acute pancreatitis treatment.

R285.5

泰安市科技发展计划（引导计划）项目（2021NS421，2022NS152）