目的 采用超高效液相-四級(jí)桿-靜電場(chǎng)軌道阱高分辨質(zhì)譜（UPLC-Q-Exactive Orbitrap-MS）聯(lián)合網(wǎng)絡(luò)藥理學(xué)及分子對(duì)接技術(shù)，探究三金片治療間質(zhì)性膀胱炎的物質(zhì)基礎(chǔ)和作用機(jī)制。方法 通過UPLC-Q-Exactive Orbitrap-MS采集數(shù)據(jù)，結(jié)合建立的數(shù)據(jù)庫(kù)表征三金片化學(xué)成分，依據(jù)SwissADME數(shù)據(jù)庫(kù)的類藥性≥3個(gè)“Yes”的評(píng)判標(biāo)準(zhǔn)，篩選活性成分；通過中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫(kù)與分析平臺(tái)（TCMSP）、SwissTargetPrediction數(shù)據(jù)庫(kù)查找活性成分靶點(diǎn)；通過基因數(shù)據(jù)庫(kù)篩選間質(zhì)性膀胱炎靶點(diǎn)。藥物靶點(diǎn)與疾病靶點(diǎn)取交集，輸入String數(shù)據(jù)庫(kù)和Cytoscape 3.7.2軟件構(gòu)建蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)，篩選出核心靶點(diǎn)。利用DAVID數(shù)據(jù)庫(kù)對(duì)核心靶點(diǎn)進(jìn)行基因本體（GO）功能注釋和京都基因與基因組百科全書（KEGG）通路富集分析；通過分子對(duì)接實(shí)驗(yàn)驗(yàn)證活性成分與核心靶點(diǎn)的結(jié)合能力，并結(jié)合分子動(dòng)力學(xué)模擬評(píng)估關(guān)鍵復(fù)合物的結(jié)合穩(wěn)定性。結(jié)果 從三金片中鑒定出51種化學(xué)成分，篩選活性成分39個(gè)；活性成分靶點(diǎn)與間質(zhì)性膀胱炎靶點(diǎn)交集165個(gè)；GO功能注釋獲得1 205條條目，KEGG通路富集分析得到190條相關(guān)信號(hào)通路，涵蓋炎癥調(diào)控、免疫應(yīng)答等關(guān)鍵生物學(xué)過程。分子對(duì)接結(jié)果顯示，木犀草素、異鼠李素、山柰酚、白藜蘆醇及柚皮苷查爾酮等活性成分，與TNF、AKT1、IL6、TP53、INS等核心靶點(diǎn)蛋白均具有良好的結(jié)合活性；分子動(dòng)力學(xué)模擬進(jìn)一步驗(yàn)證，結(jié)合能最優(yōu)的木犀草素-TNF復(fù)合物結(jié)合能為-184.22 kJ·mol^-1，表明二者結(jié)合具有高度穩(wěn)定性。結(jié)論 初步闡明三金片通過“多成分、多靶點(diǎn)、多通路”的整合調(diào)節(jié)模式治療間質(zhì)性膀胱炎的特點(diǎn)，其核心活性成分可能通過調(diào)控炎癥相關(guān)靶點(diǎn)及通路發(fā)揮作用，為后續(xù)三金片藥效機(jī)制的深入研究及臨床應(yīng)用拓展提供參考。

Objective To explore the material basis and mechanism of action of Sanjin Tablets in the treatment of interstitial cystitis by using ultra-performance liquid chromatography-quadrupole-Orbitrap high-resolution mass spectrometry（UPLC-Q-Exactive Orbitrap-MS） combined with network pharmacology and molecular docking technology. Methods Data were collected by UPLC-QExactive Orbitrap-MS and the chemical components of Sanjin Tablets were characterized based on the established database. Active components were screened according to the criterion of ≥ 3 “Yes” in drug-likeness from the SwissADME database. The target proteins of active components were identified through traditional Chinese medicine systems pharmacology database and analysis platform（TCMSP） and SwissTargetPrediction databases, and the target proteins of interstitial cystitis were screened through the gene database. The intersection of drug targets and disease targets was input into the String database and Cytoscape 3.7.2 software to construct a protein-protein interaction（PPI） network and screen out core targets. The core targets were subjected to gene ontology（GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis using the DAVID database. Molecular docking experiments were conducted to verify the binding ability of active components to core targets, and molecular dynamics simulations were used to evaluate the binding stability of key complexes. Results Fifty-one chemical components were identified in Sanjin Tablets, and 39 active components were screened. There were 165 intersections between the target proteins of active components and those of interstitial cystitis. GO functional annotation yielded 1,205 entries, and KEGG pathway enrichment analysis identified 190 related signaling pathways, covering key biological processes such as inflammation regulation and immune response. Molecular docking results showed that active components such as luteolin, isorhamnetin, kaempferol, resveratrol, and naringenin chalcone had good binding activity with core target proteins such as TNF, AKT1, IL6, TP53, and INS. Molecular dynamics simulations further verified that the binding energy of the luteolin-TNF complex was-184.22 k J·mol^-1, indicating a highly stable binding. Conclusion This study initially clarified the characteristics of Sanjin Tablets in treating interstitial cystitis through an integrated regulation mode of “multiple components, multiple targets, and multiple pathways”. Its core active components may exert their effects by regulating inflammation-related targets and pathways, providing a reference for the in-depth study of the pharmacological mechanism and clinical application expansion of Sanjin Tablets.

R285.5

全國(guó)中藥特色技術(shù)傳承人才培訓(xùn)項(xiàng)目(國(guó)中醫(yī)藥人教函[2023]96號(hào)); 廣西中醫(yī)藥大學(xué)研究生教育創(chuàng)新計(jì)劃項(xiàng)目(YCSY2025093)