Abstract: To investigate the anti ⁃ inflammatory effects of the combination of Houttuynia cordata，Rubia cordifolia，and Isatis indigotica on macrophage RAW264.7 cells through network pharmacology，molecular docking， and experimental verification methods. The active components and related target sites of Houttuynia cordata，Rubia cordifolia，and Isatis indigotica were identified using TCMSP，ETCM，UniProt，and SwissTargetPrediction databases；component⁃inflammation target network diagram was drawn using Cytoscape 3.6.1 software，and GO function and KEGG pathway enrichment analyses were performed；molecular docking was conducted using AutoDock software. The mRNA secretion of TNF⁃α，IL1⁃β，and IL⁃6 in macrophage RAW264.7 cells was detected by real⁃time fluorescent quantitative PCR to evaluate the anti ⁃ inflammatory effect of the herbal mixture. The anti ⁃ inflammatory effect of Houttuynia cordata，Rubia cordifolia，and Isatis indigotica combination was detected by mouse ear swelling test. A total of 702 active components were obtained for Houttuynia cordata，Rubia cordifolia，and Isatis indigotica， including scutellarin，corindone，and kaempferol，etc.；675 drug⁃inflammation intersection targets were obtained， including IL6，IL1β，and serine/threonine protein kinase（AKT1），etc.；GO function enrichment analysis showed 186，36，and 86 entries；KEGG result analysis yielded 189 pathways. Molecular docking results showed that the active ingredients of the drugs had good binding ability with the main targets. In vitro experiments demonstrated that the three herbal medicines could effectively inhibit the caused by Lipopolysaccharide（LPS），with pathological increases of inflammatory factors such as IL⁃6，TNF⁃α，and IL⁃1β. The combination of Houttuynia cordata，Rubia cordifolia，and Isatis indigotica inhibited the ear swelling of mice caused by dexamethasone at 47.61%，which was significantly different from the model group，indicating that the herbal mixture can exert anti⁃inflammatory effects. The good anti⁃inflammatory effect of the combination of Houttuynia cordata，Rubia cordifolia，and Isatis indigotica is demonstrated by network pharmacology analysis and experimental results，suggesting that it exerts anti⁃inflammatory effects through multiple components，multiple pathways，and multiple targets.

Key words: Inflammation, Traditional Chinese Medicine, Network Pharmacology, Signal Pathway, Molecular
Docking

摘要： 通过网络药理学、分子对接和试验验证的方法探讨鱼腥草、紫草和板蓝根合剂抗炎作用。利用TCMSP、ETCM、UniProt和SwissTargetPrediction数据库查找鱼腥草、紫草和板蓝根的活性成分和相关作用靶点；利用OMIM、TTD和Genecard数据库获取炎症的疾病靶点；采用Cyto?scape3.6.1软件绘制药物?活性成分?炎症靶点网络图，并进行GO功能和KEGG通路富集分析，应用AutoDock软件进行分子对接。通过实时荧光定量pcr方法来检测中药合剂对巨噬细胞RAW264.7中TNF?α、IL1?β、IL?6的mRNA的含量。通过小鼠耳廓肿胀试验检测鱼腥草、紫草和板蓝根合剂的抗炎作用。通过网络药理学分析鱼腥草、紫草和板蓝根共获得702个活性成分，包括紫草素、青黛酮和山柰酚等；获得675个药物与炎症交集靶点，包括IL6、IL1β和丝氨酸/苏氨酸蛋白激酶（AKT1）等；GO功能富集结果显示，获得186、36和86个条目，KEGG结果分析得到189条通路。分子对接结果显示，药物的活性有效成分与主要靶点具有良好的结合能力。体外试验表明三味中药可有效抑制脂多糖（LPS）所造成的IL?6、TNF?α和IL?1β等炎症因子病理性升高。鱼腥草、紫草和板蓝根合剂对二甲苯所致小鼠耳廓肿胀抑制率为47.61%与模型组差异显著，说明中药合剂可以发挥抗炎作用。鱼腥草、紫草和板蓝根合剂较好的抗炎效果，通过网络药理学分析和试验结果说明是通过多成分、多通路、多靶点发挥抗炎作用。

关键词: 炎症, 中药, 网络药理学, 信号通路, 分子对接