目的 构建益母草Leonurus japonicus化学成分-疾病靶点-代谢信号通路网络，探究益母草多成分-多靶点-多通路治疗寒凝血瘀型痛经的作用机制。方法 利用TCMSP数据库与Swiss Target Prediction服务器获取益母草的化学成分与作用靶点，结合DrugBank、DisGeNET、TTD等数据库获取益母草治疗寒凝血瘀型痛经的作用靶点；通过DAVID网站对作用靶点进行GO功能富集分析和KEGG通路分析，采用Cytoscape 3.6.0构建益母草活性成分-痛经作用靶点-代谢信号通路网络图，探讨益母草治疗寒凝血瘀型痛经的作用机制；并进一步采用Western blotting实验验证益母草对寒凝血瘀型痛经模型大鼠子宫组织中花生四烯酸代谢通路环氧化酶1（PTGS1）、PTGS2蛋白表达的影响。结果 获得益母草潜在活性成分共8个，与痛经相关疾病靶点22个，关键靶点涉及PTGS1、PTGS2、ALOX5、PLAG2A、AKR1C3等。GO功能富集分析共得到GO条目71个（P<0.05），其中生物过程（BP）条目46个，细胞组成（CC）条目4个，分子功能（MF）条目21个。益母草治疗痛经可能作用靶标22个，KEGG通路富集筛选得到7条信号通路（P<0.05），涉及花生四烯酸代谢、亚油酸代谢、甾类激素生物合成等；益母草可显著提高模型大鼠子宫中PTGS1、PTGS2蛋白表达水平（P<0.05），验证了网络药理学的部分预测结果。结论 益母草可能通过作用于花生四烯酸代谢通路起到治疗寒凝血瘀型痛经的作用。

Objective To establish a chemical constitution-disease target-metabolic signaling pathway network of Leonurus japonicus, and explore the mechanism of multi-component, multi-target and multi-pathway of L. japonicus for the treatment of dysmenorrhea caused by cold coagulation and blood stasis. Methods TCMSP database and Swiss Target Prediction server were used to obtain the chemical components and action targets of L. japonicus. Combined with DrugBank, DisGeNET, TTD and other databases, the action targets of L. japonicus for treating dysmenorrhea symptoms wtih cold coagulation and blood stasis were obtained. Through DAVID's website, GO function enrichment analysis and KEGG pathway analysis were conducted on the targets, and cytoscape 3.6.0 was used to construct the network diagram of the active component-dysmenorrhea target-metabolic signaling pathway of L. japonicus, to explore the mechanism of action of L. japonicus in treating dysmenorrhea with cold coagulation and blood stasis. Furthermore, Western blotting experiments were conducted to verify the effect of L. japonicus on the expression of PTGS1 and PTGS2 proteins in the uterine tissues of the model rats with cold coagulation and blood stasis dysmenorrhea. Results Eight potential active ingredients of L. japonicus were obtained, including 22 dysmenorrhea related disease targets, main targets PTGS1, PTGS2, ALOX5, PLAG2A, AKR1C3, etc. A total of 71 GO items were obtained by GO functional enrichment analysis (P<0.05), including 46 biological process (BP) items, four cell component (CC) items, and 21 molecular function (MF) items. There were 22 possible targets for the treatment of dysmenorrhea, and seven signaling pathways were obtained through KEGG pathway enrichment and screening (P<0.05), involving arachidonic acid metabolism, linoleic acid metabolism, steroid hormone biosynthesis, etc. L. japonicus significantly increased the protein expression levels of PTGS1 and PTGS2 in the uterus of the model rats (P<0.05), which confirmed some of the predicted results of network pharmacology. Conclusion L. japonicus may treat dysmenorrhea with cold coagulation and blood stasis by acting on arachidonic acid metabolism pathway.

R285.5

国家自然科学基金资助项目：组分配伍四逆汤对甲减肾损伤的保护作用及其作用机制研究（81373546）