目的 基于II型胶原诱导的类风湿性关节炎（rheumatoid arthritis，RA）大鼠模型和网络药理学方法探讨痹祺胶囊的药效作用及作用机制。方法 SD大鼠采用II型胶原蛋白造模，造模成功大鼠随机分为模型组，痹祺胶囊（0.05、0.10、0.20、0.40 g/kg）组及泼尼松（10 mg/kg）组和雷公藤总苷（10 mg/kg）组，连续给药15 d。持续监测大鼠体质量、足趾肿胀度和关节炎评分，苏木素-伊红染色考察大鼠踝关节病理变化，ELISA法测定血清细胞因子含量，初步评价痹祺胶囊治疗效果。选择痹祺胶囊中39个化合物为研究对象，依据反向药效团匹配方法和TCMSP、Uniprot等数据库预测化合物作用靶点，与通过OMIM、DisGeNet等数据库收集的RA相关靶点相互交互，将交互靶点借助Omicsbean、STRING等数据库平台对获得靶点进行基因本体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，利用Cytoscape软件构建“药材-化合物-靶点-通路-功效-疾病”网络，预测其可能的作用机制。结果 与对照组比较，模型组大鼠活动、饮食减少，足趾肿胀、关节呈急性炎症表现，关节炎评分显著增加（P＜0.01），血清中类风湿因子（rheumatoid factor，RF）、白细胞介素-17（interleukin-17，IL-17）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、IL-1β和γ-干扰素（interferon-γ，IFN-γ）水平均显著增加（P＜0.05、0.01、0.001），IL-10水平降低，脾脏、胸腺指数显著增加（P＜0.01、0.001），病理显示膝关节可见滑膜细胞增生、炎性细胞浸润，骨与软骨被侵蚀。与模型组比较，痹祺胶囊组大鼠踝关节病变情况得到不同程度改善，具体可见足肿胀度、关节炎评分显著降低（P＜0.05、0.01），血清中RF、IL-17、TNF-α、IL-1β、IFN-γ水平显著降低（P＜0.05、0.01、0.001），IL-10水平增高，脾脏、胸腺指数降低（P＜0.05、0.01），踝关节骨、软骨组织损伤减轻，炎性细胞浸润、滑膜结缔组织增生减少。网络药理学预测发现，痹祺胶囊39个成分与RA交集靶点共371个。蛋白质-蛋白质相互作用网络分析结果显示，IL-6、白蛋白、TNF、血管内皮生长因子A、基质金属蛋白酶9、趋化因子配体8等124个靶点可能是痹祺胶囊治疗RA的关键靶点。KEGG通路富集分析获得IL-17信号通路、Toll样受体信号通路、Th1和Th2细胞分化、VEGF信号通路、瞬时受体电位通道的炎症介质调节、Wnt信号通路、血小板激活等80条信号通路，与免疫调控、抗炎、抑制血管翳生成、成骨/破骨细胞平衡等过程相关。对网络进行分析发现痹祺胶囊治疗RA具有多成分、多靶点、多途径的特点，其中单体成分士的宁、三七皂苷R₁、人参皂苷Rg₁、丹参酮I、丹参酮II_A、丹酚酸B、异甘草素、迷迭香酸、阿魏酸等可能为痹祺胶囊的关键药效物质基础。结论 痹祺胶囊对II型胶原诱导的RA大鼠模型治疗效果较好，可能通过抗炎、免疫调节、血管生成、骨形成/侵蚀平衡等发挥治疗作用，体现了痹祺胶囊治疗RA多成分、多靶点、多通路的特点。

Objective To explore the pharmacodynamic effect and potential mechanism of Biqi Capsule (痹祺胶囊) based on type Ⅱ collagen-induced rheumatoid arthritis (RA) rat model and network pharmacology. Methods SD rats were modeled with type II collagen, and the successful rats were randomly divided into model group, Biqi Capsule (0.05, 0.10, 0.20, 0.40 g/kg) groups, prednison (10 mg/kg) group and Trjpterygium glycosides (10 mg/kg) group. Rats were continuously ig for 15 d, body weight, foot swelling and arthritis score of rats were continuously monitored. Hematoxylin-eosin staining was used to observe the pathological changes of ankle joint in rats, and the content of cytokines in serum was determined by ELISA, so as to preliminarily evaluate the therapeutic effect of Biqi Capsule. A total of 39 compounds in Biqi Capsule were selected as the research objects, and the targets of the compounds were predicted according to the reverse pharmacophore matching method and databases such as TCMSP and Uniprot. With the help of database platforms such as Omicsbean, STRING, etc., the interactive targets were analyzed by gene ontology function and Kyoto encyclopedia of genes and genomes (KEGG) pathway by interacting with RA-related targets collected through OMIM, DisGeNet and GeneCards, and “medicinal material-compound-target-pathway-efficacy-disease” network was constructed by Cytoscape software to predict its possible mechanism. Results Compared with control group, the activity and diet of rats in model group were reduced, the toes were swollen, the joints showed acute inflammation, the arthritis score was significantly increased (P < 0.01), and the levels of rheumatoid factor (RF), interleukin-17 (IL-17), tumor necrosis factor-α (TNF-α), IL-1β and interferon-γ (IFN-γ) in serum were significantly increased (P < 0.05, 0.01, 0.001), while the level of IL-10 was decreased, indexes of spleen and thymus were increased (P < 0.01, 0.001). Pathology showed that synovial cells were proliferated, inflammatory cells were infiltrated and bone and cartilage were eroded. Compared with model group, the pathological changes of ankle joint in Biqi Capsule group were improved in different degrees, and the foot swelling and arthritis score were significantly reduced (P < 0.05, 0.01), and the levels of RF, IL-17, TNF-α, IL-1β and IFN-γ in serum were significantly reduced (P < 0.05, 0.01, 0.001), IL-10 level was increased, indexes of spleen and thymus were decreased (P < 0.05, 0.01). The injury of ankle bone and cartilage tissue were reduced, inflammatory cell infiltration and synovial connective tissue proliferation were reduced. According to the prediction of network pharmacology, there were 371 targets where 39 components of Biqi Capsule intersected with RA. Protein-protein interaction network analysis showed that 124 targets such as IL-6, albumin, TNF, vascular endothelial growth factor A, matrix metalloproteinase 9 and chemokine ligand 8 may be the key targets of Biqi Capsule in treatment of RA. KEGG pathway enrichment analysis obtained 80 signal pathways, including IL-17 signaling pathway, Toll-like receptor signaling pathway, Th1 and Th2 cell differentiation, VEGF signaling pathway, inflammatory mediator regulation of transient receptor potential (TRP) channels, Wnt signaling pathway, platelet activation, etc., which were related to immune regulation, anti-inflammation, inhibition of pannus formation, osteogenesis/osteoclast balance and other processes. The analysis of the network showed that Biqi Capsule had the characteristics of multi-component, multi-target and multi-channel, among which the monomer components of strychnine, notoginseng saponin R₁, ginsenoside Rg₁, tanshinone I, tanshinone II_A, salvianolic acid B, isoliquiritigenin, rosmarinic acid and ferulic acid may be the key pharmacodynamic substance basis of Biqi Capsule. Conclusion Biqi Capsule has a good therapeutic effect on type II collagen-induced RA rat model, which may play a therapeutic role through anti-inflammation, immunomodulation, angiogenesis, bone formation/erosion balance, etc., reflecting the characteristics of Biqi Capsule in treating RA with multiple components, multiple targets and multiple channels.

R285.5

国家自然科学基金重点项目（U21A20406）