目的 运用血清代谢组学、网络药理学及分子对接技术探讨“黄芪-玉竹-灵芝”（HYL）配伍治疗慢性支气管炎（CB）的作用机制。方法 采用烟熏法建立SD大鼠CB模型，将模型成功大鼠随机分为模型组、阳性药（盐酸氨溴索分散片0.92 mg·kg^-1）组和HYL高、中、低剂量（10.71、3.57、1.19 g·kg^-1）组，运用酶联免疫吸附法（ELISA）测定大鼠血清中炎症因子白细胞介素8（IL-8），肿瘤坏死因子（TNF-α），一氧化氮（NO）、肺组织中P物质（SP）的表达水平。运用UPLC-Q/TOF-MS/MS技术对大鼠血清代谢谱进行分析，筛选差异代谢物及相关代谢通路；采用网络药理学，构建HYL治疗CB的“成分-靶点-通路-疾病”图。采用分子对接法验证活性成分与核心靶点的相互作用。将血清代谢组学与网络药理学联合分析，构建“靶点-代谢途径-代谢物”网络图。结果 与对照组比较，模型组大鼠血清中IL-8、TNF-α及SP水平显著降低，血清中NO水平升高；通过代谢组学技术，从血清中筛选出20个差异代谢物，与模型组相比，HYL能回调这些差异代谢物水平。筛选出5条关键代谢途径（细胞色素P450、叶酸合成、鞘脂代谢、类固醇激素生物合成、戊糖和葡萄糖醛酸相互转化）。网络药理学分析表明，HYL治疗CB主要作用于PIK3CA、MAPK3、MAPK1等靶点蛋白，与癌症通路、脂质与动脉粥样硬化等信号通路密切相关。整合分析显示细胞色素P450和类固醇激素生物合成代谢是HYL治疗CB的关键代谢途径。结论 HYL可通过调节血清代谢物调控炎症相关核心靶点基因，降低炎症水平，缓解机体代谢紊乱发挥治疗CB的作用。

Objective To investigate the mechanism of action of "Astragali Radix-Polygonati Odorati Rhizoma-Ganoderma" (HYL) in the treatment of chronic bronchitis (CB) using serum metabolomics, network pharmacology and molecular docking techniques. Methods A chronic bronchitis model was established in SD rats by smoking method, and the successful rats were randomly divided into the model group, the positive drug group (Ambroxol Hydrochloride Dispersible Tablets 0.92 mg·kg^-1), the HYL high-,mid-, low-doses (10.71, 3.57, 1.19 g·kg^-1) groups, and the enzyme-linked immunosorbent assay (ELISA) was used to determine the inflammatory factors interleukin 8 (IL-8), tumor necrosis factor (TNF-α), and nitric oxide (NO) in the serum of the rats, substance P (SP) expression levels in lung tissues. The metabolic profiles of rat serum were analyzed by UPLC-Q/TOF-MS/MS, and the differential metabolites and related metabolic pathways were screened. The "ingredient-target-pathway-disease" diagram of HYL for CB treatment was constructed by network pharmacology. The molecular docking method was used to verify the interaction between the active ingredients and the core targets. Serum metabolomics and network pharmacology were combined to construct a "target-metabolic pathway-metabolite" network diagram. Results HYL significantly reduced the levels of IL-8, TNF-α and SP, and increased the level of NO in the serum of rats. Twenty differential metabolites were screened out from serum by metabolomics, and HYL was able to regulate the levels of these differential metabolites compared with those in the model group. Five key metabolic pathways (cytochrome P450, folate synthesis, sphingolipid metabolism, steroid hormone biosynthesis, and pentose and glucuronide interconversion) were screened. Network pharmacological analyses showed that HYL therapeutic CB mainly acted on target proteins such as PIK3CA, MAPK3 and MAPK1, which are closely related to signaling pathways such as cancer pathways, lipids and atherosclerosis. Integrative analysis showed that cytochrome P450 and steroid hormone biosynthesis metabolism were the key metabolic pathways in HYL treatment of CB. Conclusion HYL can play a therapeutic role in CB treatment by regulating serum metabolites to modulate the core target genes related to inflammation, reducing inflammation level and alleviating metabolic disorders in the body.

R285.5

黑龙江中医药大学优秀创新人才(领军人才)基金项目(2018RCL13)