目的 根据正源方具有缓解化疗后患者白细胞降低的药理作用，分析正源方的化学成分，并进行网络药理学研究和动物实验验证。方法 使用UPLC-TOF/MS技术检测正源方的主要化学成分，在数据库中检索主要化学成分作用靶点和白细胞减少症相关靶点，用韦恩图得到交集靶点。运用Cytoscape 3.7.2软件构建交集靶点的蛋白相互作用（protein-protein interaction，PPI）网络图和“中药-成分-靶点”网络图，筛选出度值高的关键成分和靶点。将交集靶点上传R软件进行基因本体（gene ontology，GO）功能富集分析和京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）富集分析。将SD雄性大鼠随机分为空白组、模型组、阳性药组［地榆升白片0.11 g/（kg·d）］、正源方高剂量组［2.73 g/（kg·d）］、中剂量组［1.37 g/（kg·d）］、低剂量组［0.68 g/（kg·d）］，除空白组外，各组大鼠ip环磷酰胺［40 mg/（kg·d）］4 d诱导白细胞减少症大鼠模型，测定白细胞数、脏器指数，ELISA试剂盒检测肿瘤坏死因子-α（tumor necrosis factor，TNF-α）、白细胞介素-6（interleukin-6，IL-6）的含量。结果 使用UPLC-TOF/MS技术共测得24个化学成分。网络药理学结果显示，正源方的活性成分作用于377个靶点，检索到疾病相关靶点1952个，交集靶点124个。筛选得出人参皂苷Rg₁、阿魏酸、人参皂苷Rb₁、人参皂苷Rg₂、人参皂苷Ro、山柰酚等核心成分；TNF和IL-6等共20个重要靶点，富集到的关键通路有IL-17信号通路（IL-17 signaling pathway）等。动物实验验证结果显示，与模型组相比，正源方高、中剂量给药组的大鼠白细胞计数和胸腺指数显著升高，血清中TNF-α、IL-6水平显著降低（P<0.05），能够减轻白细胞减少症大鼠脾脏和胸腺组织的病理损伤。结论 正源方治疗白细胞减少症的物质基础是以人参皂苷为主的化学成分，相关作用机制具有多靶点、多通路的特点。

Objective According to the pharmacological effects of alleviating patient's leukopenia after chemotherapy of Zhengyuan Prescription (正源方), the chemical components of Zhengyuan Prescription were analyzed, network pharmacology study and animal experiment verification was conducted. Methods The main chemical components of Zhengyuan Prescription was detected by UPLC-TOF/MS technology, the action targets of main chemical components and the targets of leukopenia were searched through the databases. Then the Venn diagram was used to get the intersection targets. The protein-protein interaction network of intersection targets and "herb-ingredient-target" network map were constructed by Cytoscape 3.7.2 software, meanwhile, the key components and targets with high degrees were screened out. R software was used to perform gene ontology function enrichment analysis and Kyoto encyclopedia of genes and genomes enrichment analysis. The healthy male SD rats were divided into the blank control group, model control group, positive drug group 0.11 g/(kg·d) (Diyu Shengbai Tablets), Zhengyuan Prescription high-dose[2.73 g/(kg·d)] medium-dose[1.37 g/(kg·d)], and low-dose[0.68 g/(kg·d)] groups. Cyclophosphamide[40 mg/(kg·d)] was given by intraperitoneal injection for 4 d to induce leukopenia rat model. The number of white blood cell was counted, and organ index was detected. Tumor necrosis factor (TNF) and interleukin-6 (IL-6) contents were detected with ELISA kit. Results Twenty-four main chemical components of Zhengyuan Prescription were obtained by UPLC-TOF/MS technology. Network pharmacology results showed that there were many active components in Zhengyuan Prescription, which acted on 377 targets, 1952 disease-related targets were retrieved, and 124 targets were intersected. The core components were screened such as ginsenoside Rg₁, ferulic acid, ginsenoside Rb₁ ginsenoside Rg₂, ginsenoside Ro, kaempferol; The key targets were screened such as IL-6 and TNF, the important pathways including IL-17 signaling pathway and so on. The animal experiments results showed that:Compared with the model group, the high and medium doses of Zhengyuan Prescription administration groups significantly increased the number of white blood cell and thymus index of rats, significantly reduced the elevated TNF-α and IL-6 levels in the serum (P < 0.05), and it can reduce the pathological damage of spleen and thymus tissues in rats with leukopenia. Conclusion The material basis of Zhengyuan Prescription for treating leukopenia is based on ginsenoside-based chemical components, and the related mechanism of action has the characteristics of multiple targets and multiple pathways.

R283.5

国家重大科技专项“重大新药创制”项目（2019ZX09301-133）；陕西省科技厅重点产业创新链（群）项目（2020ZDLSF05-08）；现代农业产业技术体系建设专项资金（CARS-21）