目的 基于网络药理学整合体内实验方法探究脉络舒通丸抗血栓性浅静脉炎（superficial thrombophlebitis，STP）的作用机制。方法 借助网络药理学方法，从药物及疾病相关数据库筛选脉络舒通丸的成分作用靶点及STP疾病相关靶点，根据拓扑特征值筛选关键靶点后，进行基因本体（gene ontology，GO）功能及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。将日本大耳白兔随机分为对照组、模型组、地奥司明（0.168 g/kg）组和脉络舒通丸低、中、高剂量（0.56、1.68、5.04 g/kg）组。除对照组外，其余各组均采用耳缘iv 20%甘露醇建立STP兔模型。造模同时各给药组连续ig给药14 d。采用苏木素-伊红（HE）染色法检测各组兔耳组织病理变化；ELISA检测各组血清中白细胞介素-1β（interleukin-1β，IL-1β）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）和IL-6水平；Western blotting检测耳组织蛋白激酶B（protein kinase B，Akt）、磷脂酰肌醇-3-激酶（phosphatidylin-ositol-3-kinase，PI3K）、Janus激酶2（Janus kinase 2，JAK2）和信号传导与转录激活因子3（signal transducer and activator of transcription 3，STAT3）蛋白表达及磷酸化水平。结果 共筛选得到脉络舒通丸活性成分作用靶点1112个，STP疾病相关靶点837个，二者共同靶点84个，根据拓扑特征值筛选出核心靶点31个，富集分析发现核心靶点主要与平滑肌细胞增殖的正调控、炎症反应的正调控、凋亡过程的调控等生物过程，以及JAK/STAT信号通路、PI3K/Akt信号通路有关。体内动物实验结果显示，脉络舒通丸组兔耳组织病理损伤显著改善，血清中IL-1β、TNF-α和IL-6水平显著降低（P＜0.05、0.01），耳组织Akt、PI3K、JAK2和STAT3蛋白的磷酸化水平均显著下调（P＜0.05、0.01）。结论 脉络舒通丸可能通过下调PI3K/Akt和JAK2/STAT3信号通路相关蛋白表达，抑制炎症和血栓的形成，从而发挥抗STP的作用。

Objective To explore the mechanism of Mailuo Shutong Pills (脉络舒通丸, MLSTW) in treatment of superficial thrombophlebitis (STP) based on combination of network pharmacology and animal experimental validation in vivo. Methods Drug targets of MLSTW and targets related to STP were screened from disease and drug related database, and key targets were screened according to topological eigenvalues. Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed. The Japanese white rabbits were randomly divided into control group, model group, diosmin (0.168 g/kg) group and MLSTW low-, medium-, high-dose (0.56, 1.68, 5.04 g/kg) groups. Except for control group, the rabbit model of STP was established by injecting 20% mannitol into bilateral auricular vein in other groups. At the same time, the treated groups were ig drugs for 14 d. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of rabbit ear tissues; ELISA was used to detect the levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and IL-6 in serum; Western blotting was used to detect the expressions of protein kinase B (Akt), phosphatidylin-ositol-3-kinase (PI3K), Janus kinase 2 (JAK2), signal transducer and activator of transcription 3 (STAT3) proteins and phosphorylation levels in rabbit ear tissues. Results A total of 1112 compounds targets of MLSTP, 837 STP disease targets, 84 common targets and 31 key targets were obtained. Enrichment analysis showed that the key targets were mainly involved in biological processes such as positive regulation of smooth muscle cell proliferation, positive regulation of inflammatory response, positive regulation of apoptotic process, as well as PI3K/Akt signaling pathway, JAK/STAT signaling pathway. In vivo experiment results showed that pathological damage of rabbit ear tissues in MLSTW group was significantly improved, levels of IL-1β, TNF-α, IL-6 in serum of rabbits were significantly decreased (P < 0.05, 0.01), phosphorylation levels of Akt, PI3K, JAK2 and STAT3 in rabbit ear tissues were significantly reduced (P < 0.05, 0.01). Conclusion MLSTP can play an anti-STP role by down-regulating the expressions of PI3K/Akt and JAK2/STAT3 signaling pathway related proteins, inhibiting inflammation and thrombosis.

R285.5

国家自然科学基金资助项目（81973557）；天津市自然科学基金重点项目（20JCZDJC00010）；释药技术与药代动力学国家重点实验室（天津药物研究院）开发课题资助（010162001）