目的 探究对香豆酸抑制炎症途径改善急性心肌缺血（acute myocardial ischemia，AMI）的保护作用和分子机制。方法 应用PubChem等数据库预测筛选对香豆酸与AMI的共同靶点，将获得的共同靶点导入STRING数据库构建蛋白相互作用网络（protein-protein interaction，PPI），并对PPI结果进行拓扑分析；利用DAVID数据库和微生信平台进行基因本体（gene ontology，GO）功能和京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析；运用分子对接技术和分子动力学模拟分析对香豆酸与Toll样受体4（Toll-like receptor 4，TLR4）共受体髓样分化因子2（myeloid differentiation 2，MD2）的结合能力和稳定性。采用冠状动脉左前降支结扎法建立AMI小鼠模型，设置假手术组、模型组、对香豆酸（100 mg/kg）组、卡维地洛（7.6 mg/kg）组及TAK-242（3 mg/kg）组，连续给药7 d，末次给药后采用超声心动评价对香豆酸对心功能的保护作用；采用苏木素-伊红（hematoxylin-eosin，HE）及Masson染色观察心肌组织病理学损伤；检测血清中肌酸激酶同工酶（creatine kinase-MB，CK-MB）活性、心肌肌钙蛋白I（cardiac troponin I，cTnI）水平和心肌组织炎症因子水平；采用Western blotting检测心肌组织TLR4/核因子-κB（nuclear factor-κB，NF-κB）炎症通路相关蛋白的表达。构建H9c2心肌细胞缺氧炎症复合损伤模型，进一步探究对香豆酸对TLR4/NF-κB炎症途径的调控机制。结果 网络药理学分析显示对香豆酸可作用于TLR4等核心靶点并调控TLR及NF-κB通路治疗AMI，分子对接结果显示对香豆酸与TLR4共受体MD2具有较好的亲和力，分子动力学模拟表明对香豆酸-MD2具有良好的稳定性。动物实验结果显示，对香豆酸能显著改善AMI小鼠心功能（P＜0.01、0.001），减轻心肌组织炎症细胞浸润，抑制胶原蛋白沉积及心肌纤维化，降低血清中CK-MB活性及cTnI水平（P＜0.01、0.001），下调心肌组织中TLR4、p-NF-κB/NF-κB、NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor thermal protein domain associated protein 3，NLRP3）、gasdermin D蛋白（gasdermin D，GSDMD）和白细胞介素-1β（interleukin-1β，IL-1β）蛋白表达以及IL-6水平（P＜0.05、0.01、0.001）。细胞实验结果显示，对香豆酸可显著抑制TLR4、p-NF-κB/NF-κB、NLRP3蛋白表达水平（P＜0.05、0.01）。结论 对香豆酸可显著改善AMI小鼠心功能、抑制心肌组织炎症反应及心肌纤维化，其作用机制与调控TLR4/NF-κB炎症途径有关。

Objective To investigate the protective effects and underlying molecular mechanisms of p-coumaric acid (p-CA) against acute myocardial ischemia (AMI) through inhibition of inflammatory signaling. Methods Common targets of p-CA and AMI were predicted and screened using databases such as PubChem. These common targets were imported into the STRING database to construct a protein-protein interaction (PPI) network, followed by topological analysis. Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed using DAVID database and bioinformatics platform. The binding affinity and stability between p-CA and myeloid differentiation 2 (MD2), a co-receptor of Toll-like receptor 4 (TLR4), were analyzed using molecular docking and molecular dynamics simulations. An AMI mouse model was established by ligating the left anterior descending coronary artery. The mice were randomly assigned into sham group, model group, p-CA (100 mg/kg) group, carvedilol (7.6 mg/kg) group and TAK-242 (3 mg/kg) group. After seven consecutive days of treatment, echocardiography was performed to evaluate cardiac function. Myocardial histopathological damage was observed using hematoxylin-eosin (HE) and Masson staining. Activity of creatine kinase-MB, level of cardiac troponin I (cTnI) in serum and levels of inflammatory factors in myocardial tissue were detected. The expressions of proteins related to TLR4/nuclear factor-κB (NF-κB) inflammatory pathway in myocardial tissue were measured by Western blotting. A hypoxia-inflammatory complex injury model in H9c2 cardiomyocytes was constructed to further evaluate the regulatory mechanism of p-CA on TLR4/NF-κB inflammatory signaling pathway. Results Network pharmacology analysis indicated that p-CA could act on core targets such as TLR4 and regulate TLR and NF-κB pathways to treat AMI. Molecular docking results demonstrated that p-CA exhibited favorable binding affinity with MD2, a co-receptor of TLR4. Molecular dynamics simulations further verified that the p-CA-MD2 complex possessed good structural stability. The results of animal experiments showed that p-CA significantly improved cardiac function in AMI mice (P < 0.01, 0.001), alleviated inflammatory cell infiltration in myocardial tissue, inhibited collagen deposition and myocardial fibrosis, and reduced activity of CK-MB and level of cTnI in serum (P < 0.01, 0.001). Furthermore, p-CA downregulated TLR4, p-NF-κB/NF-κB, NOD-like receptor thermal protein domain associated protein 3 (NLRP3), gasdermin D (GSDMD), interleukin-1β (IL-1β) protein expressions and IL-6 level in myocardial tissue (P < 0.05, 0.01, 0.001). The results of cell experiments confirmed that p-CA significantly suppressed the protein expression levels of TLR4, p-NF-κB/NF-κB and NLRP3 (P < 0.05, 0.01). Conclusion p-CA could improve cardiac function in AMI, inhibit myocardial inflammatory response and fibrosis, and its mechanism is associated with the regulation of TLR4/NF-κB inflammatory pathway.

R285.5

国家自然科学基金资助项目（82004095）；中央高校基本科研业务费专项资助项目（2023-JYB-JBQN-013，2021-JYB-XJSJJ031）；国家中医药管理局司便函（国中医药科技中药便函[2022]59号）；国家中医药管理局科技司项目（GZY-KJS-2022-057）；曹俊岭全国老药工传承工作室建设项目（国中医药人教函2024-255）；曹俊岭临方炮制中医药传统技能传承工作室（2025-JNZZS-13）