目的 運用血清代謝組學、網(wǎng)絡藥理學及分子對接技術探討“黃芪-玉竹-靈芝”（HYL）配伍治療慢性支氣管炎（CB）的作用機制。方法 采用煙熏法建立SD大鼠CB模型，將模型成功大鼠隨機分為模型組、陽性藥（鹽酸氨溴索分散片0.92 mg·kg^-1）組和HYL高、中、低劑量（10.71、3.57、1.19 g·kg^-1）組，運用酶聯(lián)免疫吸附法（ELISA）測定大鼠血清中炎癥因子白細胞介素8（IL-8），腫瘤壞死因子（TNF-α），一氧化氮（NO）、肺組織中P物質(zhì)（SP）的表達水平。運用UPLC-Q/TOF-MS/MS技術對大鼠血清代謝譜進行分析，篩選差異代謝物及相關代謝通路；采用網(wǎng)絡藥理學，構建HYL治療CB的“成分-靶點-通路-疾病”圖。采用分子對接法驗證活性成分與核心靶點的相互作用。將血清代謝組學與網(wǎng)絡藥理學聯(lián)合分析，構建“靶點-代謝途徑-代謝物”網(wǎng)絡圖。結果 與對照組比較，模型組大鼠血清中IL-8、TNF-α及SP水平顯著降低，血清中NO水平升高；通過代謝組學技術，從血清中篩選出20個差異代謝物，與模型組相比，HYL能回調(diào)這些差異代謝物水平。篩選出5條關鍵代謝途徑（細胞色素P450、葉酸合成、鞘脂代謝、類固醇激素生物合成、戊糖和葡萄糖醛酸相互轉化）。網(wǎng)絡藥理學分析表明，HYL治療CB主要作用于PIK3CA、MAPK3、MAPK1等靶點蛋白，與癌癥通路、脂質(zhì)與動脈粥樣硬化等信號通路密切相關。整合分析顯示細胞色素P450和類固醇激素生物合成代謝是HYL治療CB的關鍵代謝途徑。結論 HYL可通過調(diào)節(jié)血清代謝物調(diào)控炎癥相關核心靶點基因，降低炎癥水平，緩解機體代謝紊亂發(fā)揮治療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

黑龍江中醫(yī)藥大學優(yōu)秀創(chuàng)新人才(領軍人才)基金項目(2018RCL13)