目的 基于超高效液相色譜串聯(lián)四極桿靜電場軌道阱質(zhì)譜（UPLC-Q Exactive Orbitrap-HRMS）技術(shù)、網(wǎng)絡(luò)藥理學(xué)和分子對接探究黃連上清丸解熱抗炎的活性成分及其作用機制。方法 根據(jù)質(zhì)譜信息、對照品裂解規(guī)律及參考文獻資料對黃連上清丸中的化學(xué)成分進行指認；利用SwissADME數(shù)據(jù)庫篩選黃連上清丸中的成藥性成分，PharmMapper數(shù)據(jù)庫獲取成藥性成分相關(guān)作用靶點；利用Gene cards和Disgenet數(shù)據(jù)庫篩選解熱、炎癥相關(guān)靶點；對得到的成分作用靶點與疾病相關(guān)靶點取交集，獲得黃連上清丸發(fā)揮解熱抗炎作用的潛在靶點，采用Cytoscape 3.10.1軟件構(gòu)建“成分-靶點-疾病-復(fù)方”網(wǎng)絡(luò)，分析活性成分；將共有靶點導(dǎo)入STRING網(wǎng)站，構(gòu)建蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)，分析關(guān)鍵靶點；通過微生信網(wǎng)站對關(guān)鍵靶點進行基因本體（GO）和京都基因與基因組百科全書（KEGG）富集分析；將活性成分與關(guān)鍵靶點進行分子對接驗證；通過建立HPLC指紋圖譜，控制黃連上清丸的質(zhì)量。結(jié)果 從黃連上清丸中共鑒定144個化合物，其中黃酮類58個、生物堿類17個、環(huán)烯醚萜類7個、木脂素類4個、色原酮類8個、蒽醌類7個、香豆素類2個、皂苷類5個、有機酸類5個、酚酸類7個、苯丙素類9個、苯乙醇苷類4個、糖類2個、內(nèi)酯類1個，以及其他類8個。黃連上清丸發(fā)揮解熱抗炎作用的潛在作用靶點有218個，涉及2 036個生物過程、119個細胞組分、281個分子功能和155條信號通路。篩選得到蓮心季銨堿、甘草寧G、大黃素、氧化小檗堿、甘草素、大黃素甲醚、花椒毒酚、粘毛黃芩素Ⅲ等12個潛在活性成分，胰島素（INS）、RAC-α絲氨酸/蘇氨酸蛋白激酶（AKT1）、表皮生長因子受體（EGFR）、熱休克蛋白HSP 90-α（HSP90AA1）、基質(zhì)金屬蛋白酶-9（MMP9）等10個關(guān)鍵靶蛋白，以及Ras信號通路、叉頭框蛋白O信號通路、Rap1信號通路、絲裂原活化蛋白激酶信號通路等信號通路。12個潛在活性成分與關(guān)鍵靶點有較強的結(jié)合活性。建立的指紋圖譜可指認黃連上清丸中的8個活性成分。結(jié)論 黃連上清丸可能是通過蓮心季銨堿、甘草寧G、大黃素、氧化小檗堿、甘草素、大黃素甲醚等活性成分，作用于INS、AKT1、EGFR、HSP90AA1、MMP9等關(guān)鍵靶點，通過Ras信號通路、叉頭框蛋白O信號通路、Rap1信號通路、絲裂原活化蛋白激酶信號通路等信號通路，發(fā)揮解熱抗炎作用。

Objective To investigate the active components and its mechanism for the effect on antipyretic and anti-inflammatory in Huanglian Shangqing Pills using UPLC-Q Exactive Orbitrap-HRMS, network pharmacology and molecular docking.Methods According to the mass spectrometry information, the cleavage law of the reference substance and the reference literatures, the chemical compositions in Huanglian Shangqin Pills were identified; SwisSADME database was used to screen the druggable components in Huanglian Shangqing Pills, and the PharmMapper database was used to obtain the relevant targets of the druggable components; Antipyretic and inflammation-related targets was screened by Gene cards and Disgenet databases; The intersections between the obtained component targets and disease-related targets were used to obtain the potential targets of Huanglian Shangqing Pills to exert antipyretic and anti-inflammatory effects, and the "component-target-disease-compound" network was constructed by Cytoscape 3.10.1 software to analyze the active ingredients; The common targets were imported into the String website for protein-protein interaction analysis to search for key targets; GO function and KEGG pathway enrichment analysis of key targets were carried out through bioinformatics website; Molecular docking was used to validate the active ingredients and key targets; The quality of Huanglian Shangqing Pills was controlled by establishing HPLC fingerprints. Results A total of 144 compounds were identified in Huanglian Shangqing Pills, including 58 flavonoids, 17 alkaloids, 7 iridoid terpenoids, 4 lignans, 8 chromogenone, 7 anthraquinones, 2 coumarins, 5 saponins, 5 organic acids, 7 phenolic acids, 9 phenylpropanoids, 4 phenylethanol glycosides, 2 sugars, 1 lacone, and 8 other substances. There were 218 potential targets for antipyretic and anti-inflammatory effects, involving 2036 biological processes, 119 cellular components, 281 molecular functions and 155 signaling pathways. Twelve potential active ingredients including lotusine, gancaonin G, emodin, berlambine, glycyrrhizin, physcion, xanthotoxol, ganhuangenin and others, 10 key target proteins such as INS, AKT1, EGFR, HSP90AA1, MMP9 and so on, and signaling pathways such as Ras signaling pathway, FoxO signaling pathway, Rap1 signaling pathway, and MAPK signaling pathway were screened. The 12 potential active ingredients have strong binding activity to key targets. The established fingerprint identified eight active ingredients in Huanglian Shangqing Pills. Conclusion Huanglian Shangqing Pills may act on key targets such as INS, AKT1, EGFR, HSP90AA1, MMP9, etc. through active components such as lotusine, gancaonin G, emodin, berlambine, glycyrin, physcion, etc., and play the role through the signaling pathways such as Ras signaling pathway, FoxO signaling pathway, Rap1 signaling pathway, and MAPK signaling pathway, etc.

R285.5

國家自然科學(xué)基金青年項目（82301725）;中國博士后科學(xué)基金第73批面上資助項目（2023M732155）;山西省科技廳基礎(chǔ)研究項目（20210302124586）;山西省重點研發(fā)項目（202102130501023）;太原市科技局“雙百攻關(guān)行動”首批關(guān)鍵核心技術(shù)攻關(guān)“揭榜掛帥”項目（2024TYJB0147）;山西省技術(shù)創(chuàng)新中心項目（202104010911006）;山西省教育廳教改項目（J20230518）