目的 利用网络药理学方法，探究暖宫七味丸治疗痛经的作用机制并采用痛经模型大鼠对其进行实验验证。方法 使用中药系统药理学数据库与分析平台（TCMSP）、中医药整合药理学研究平台（TCMIP）和网络药理学在线数据库（TCMID）结合文献报道确定暖宫七味丸的活性成分，通过Swiss Target Prediction在线平台得到活性成分对应的靶点。利用DisGeNET数据库和GeneCard数据库确定原发性痛经的相关靶点，手动筛选化合物-疾病的共同靶点，将潜在共同靶点蛋白上传至Biogrid平台，获取药物-疾病靶蛋白相互作用（PPI）网络，利用Cytoscape 3.7.2软件对PPI网络进行网络图优化和拓扑学分析。利用DAVID 6.8数据库对潜在靶点进行京都基因与基因组百科全书（KEGG）通路富集分析及基因本体论（GO）注释分析。动物实验验证采用缩宫素致痛经大鼠模型观察暖宫七味丸高、中、低剂量（2.0、1.0、0.5 g·kg^-1）的镇痛作用，采用Western blotting方法观察暖宫七味丸高、低剂量（2.0、0.5 g·kg^-1）对痛经模型大鼠子宫组织环氧合酶-2（COX-2）蛋白表达的影响，并测定各组大鼠子宫组织中前列腺素F2α（PGF2α）水平。通过大鼠离体子宫肌张力实验观察暖宫七味丸水溶液（22.5、11.25 mg·mL^-1）对大鼠子宫收缩幅度和收缩频率的影响。结果 网络药理学预测结果显示，暖宫七味丸治疗痛经的潜在靶点有68个，PPI核心靶点有IL-6、PTGS2、TNF和TP53等。涉及到的通路有炎症相关的信号通路、癌症通路和雌激素信号通路等，涉及到的生物功能有炎症反应、一氧化氮的生物调节过程和对激素的反应和对血管的调节功能等。动物实验结果显示，与模型组比较，暖宫七味丸高、中、低剂量组和阳性药组大鼠的扭体反应次数显著减少（P＜0.01、0.001），同时扭体反应的潜伏期均明显延长（P＜0.01、0.001）；暖宫七味丸高、低剂量组痛经模型大鼠子宫组织中过氧化物合成酶2（PTGS2）相关蛋白COX-2表达显著下调（P＜0.001）），同时子宫组织中PGF2α水平显著降低（P＜0.001）。暖宫七味丸高、低剂量水提液对大鼠子宫收缩幅度和频率均有显著的抑制作用（P＜0.001）。结论 暖宫七味丸可以通过下调PTGS2靶点进而调控子宫肌张力起到治疗痛经的效果。

Objective To explore the mechanism of Nuangong Qiwei Pill in treatment of primary dysmenorrhea by network pharmacological methods and to verify the prediction results by animal experiments. Methods The active components of Nuangong Qiwei Pills were screened by using the traditional Chinese Medicine System Pharmacology database and analysis platform (TCMSP), traditional Chinese medicine integrated pharmacology research platform (TCMIP) and online pharmacology database (TCMID) combined with literature reports, and the corresponding targets of the active components were obtained through the Swiss target prediction online platform. The relevant targets of primary dysmenorrhea were obtained by using the database of DisGeNET and GeneCard, the common targets of compounds and diseases were manually screened, the potential common target proteins were uploaded to the Biogrid platform. The protein-protein interaction (PPI) network of drug-disease target was obtained, and the network diagram optimization and topology analysis of PPI network were carried out by using the software of Cytoscape 3.7.2. David 6.8 database was used to analyze the enrichment of Kyoto Encyclopedia of genes and genomes (KEGG) pathway and Gene Ontology (GO) of potential targets. The rat model of dysmenorrhea caused by oxytocin was used to observe the analgesic effect of high, medium and low doses (2.0, 1.0, 0.5 g·kg^-1) of Nuangong Qiwei Pills, and the effect of high and low doses (2.0, 0.5 g·kg^-1) of Nuangong Qiwei Pills on the expression of cyclooxygenase-2 (COX-2) protein in the uterus of dysmenorrhea rats was observed by Western blotting method, and prostaglandin F2α (PGF2α) level in the uterus of rats in each group was measured. The effects of the aqueous solution of Nuangong Qiwei Pills (22.5, 11.25 mg·mL^-1) on the amplitude and frequency of uterine contraction in rats were observed by the tension test of isolated uterine muscles in rats. Results The predicted results of network pharmacology showed that there were 68 potential targets of Nuangong Qiwei Pills in treatment of dysmenorrhea, and the core targets of PPI were IL-6, PTGS2, TNF and TP53. The pathways involved include inflammation related signaling pathways, cancer pathways and estrogen signaling pathways, and the biological functions involved include inflammatory response, biological regulation process of nitric oxide, response to hormones and regulation function of blood vessels. The results of animal experiments showed that compared with the model group, the number of writhing reaction of rats in the high, medium and low dose of Nuangong Qiwei Pills groups and positive drug groups was significantly reduced (P < 0.01, 0.001), and the incubation period of writhing reaction was significantly prolonged (P < 0.01, 0.001). The expression of PTGS2 related protein COX-2 in the uterus of dysmenorrhea model rats in the high, medium and low dose groups of Nuangong Qiwei Pills groups was significantly down regulated (P < 0.001), and the level of PGF2α in the uterus decreased significantly (P < 0.001). The high and low doses of water extract of Nuangong Qiwei Pills had significant inhibitory effects on the amplitude and frequency of uterine contraction in rats (P < 0.001). Conclusion Nuangong Qiwei Pills have therapeutic effect on dysmenorrhea by down regulating PTGS2 target and then regulating uterine muscle tension.

R285.5

国家重点研发计划（2018YFC1708203）；中央高校基本科研业务费专项（3332021064）；内蒙古自治区科技计划（2020GG0199）