目的 通過高效液相色譜-四極桿-飛行時(shí)間質(zhì)譜（HPLC-Q-TOF-MS/MS）分析兒瀉康貼膜體外透皮接收液的化學(xué)成分，結(jié)合網(wǎng)絡(luò)藥理學(xué)和分子對(duì)接技術(shù)初步預(yù)測其治療小兒腹瀉、腹痛的藥效物質(zhì)基礎(chǔ)及作用機(jī)制。方法 采用 HPLC-QTOF-MS/MS 于正、負(fù)離子模式下掃描，確認(rèn)兒瀉康貼膜透皮化學(xué)成分。選取 19 個(gè)主要活性成分為研究對(duì)象，利用 SwissTarget prediction數(shù)據(jù)庫收集化合物潛在作用靶點(diǎn)；通過GeneCards、OMIM、Drugbank數(shù)據(jù)庫獲取小兒腹瀉、腹痛相關(guān)疾病靶點(diǎn)；借助String 12.0網(wǎng)絡(luò)分析平臺(tái)，獲得活性成分與疾病交集靶點(diǎn)的蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)，從而篩選出核心靶點(diǎn)；利用 Omicsbean在線分析平臺(tái)對(duì)交集靶點(diǎn)進(jìn)行基因本體（GO）功能分析和京都基因與基因組百科全書（KEGG）通路富集分析，進(jìn)一步通過Cytoscape3.9.1軟件構(gòu)建網(wǎng)絡(luò)圖；運(yùn)用Schrödinger2020 Maestro12.4軟件將10個(gè)主要活性成分與15個(gè)核心靶點(diǎn)進(jìn)行分子對(duì)接，驗(yàn)證其結(jié)合能力。結(jié)果 兒瀉康貼膜體外透皮接收液共鑒定出90個(gè)成分，包括生物堿類、黃酮類、有機(jī)酸類、苯丙素類、三萜類、苦味素及其他類；選取的19個(gè)活性成分治療小兒腹瀉、腹痛的潛在作用靶點(diǎn)共260個(gè)，經(jīng)PPI網(wǎng)絡(luò)分析篩選出腫瘤蛋白p53（TP53）、信號(hào)傳導(dǎo)及轉(zhuǎn)錄激活蛋白3（STAT3）、表皮生長因子受體（EGFR）、絲氨酸/蘇氨酸蛋白激酶1（AKT1）等15個(gè)核心靶點(diǎn)；KEGG通路富集共得到166條通路，篩選出與疾病密切相關(guān)的通路共90條，包括磷脂酰肌醇-3-激酶-蛋白激酶 B信號(hào)通路、絲裂原活化蛋白激酶信號(hào)通路、神經(jīng)營養(yǎng)素信號(hào)通路、白細(xì)胞介素 17信號(hào)通路等重要信號(hào)通路。分子對(duì)接結(jié)果顯示，兒瀉康貼膜中10個(gè)主要活性成分與15個(gè)核心靶點(diǎn)蛋白間結(jié)合能力較強(qiáng)，能形成穩(wěn)定的復(fù)合物。結(jié)論 通過HPLC-Q-TOF-MS/MS獲得兒瀉康貼膜體外透皮成分，結(jié)合網(wǎng)絡(luò)藥理學(xué)和分子對(duì)接技術(shù)初步確定了兒瀉康貼膜發(fā)揮多成分、多靶點(diǎn)、多途徑治療小兒腹瀉、腹痛的潛在藥效物質(zhì)基礎(chǔ)及作用機(jī)制。

Objective To analyze the chemical components of the transdermal receiving solution of Erxiekang Emplastra in vitro by HPLC-Q-TOF-MS/MS, and to preliminarily predict the pharmacodynamic material basis and mechanism of its efficacy in treating children's wind-cold diarrhea by network pharmacology and molecular docking technology. Methods HPLC-Q-TOF-MS/MS was used to scan in positive and negative ion mode to confirm the transdermal chemical components of Erxiekang Emplastra. A total of 19 main active components were selected as the research objects, and the potential targets of the compounds were collected by Swiss Target prediction database. The targets of diseases related to wind-cold diarrhea in children were obtained through GeneCards, OMIM and Drugbank databases. STRING network analysis platform was used to obtain the protein-protein interaction (PPI) network of active ingredients and disease targets was obtained, so as to screen out the core targets. On-line analysis platform of Omicsbean was used to analyze the function of gene ontology (GO) and the pathway enrichment of Kyoto Gene and Genome Encyclopedia (KEGG), and then the network diagram was constructed by Cytoscape software. Schrodinger2020 Maestro12.4 software was used to perform molecular docking between 10 main active ingredients and 15 core targets to verify their binding ability. Results A total of 90 components were identified from the transdermal receiving solution of Erxiekang Emplastra in vitro, including alkaloids, flavonoids, organic acids, phenylpropanoids, triterpenoids, picrosins and others. There were 260 potential targets of 19 active ingredients in the treatment of wind-cold diarrhea in children, and 15 core targets, such as tumor protein p53 (TP53), signal transduction and transcription activating protein 3 (STAT3), epidermal growth factor receptor (EGFR) and serine/threonine protein kinase 1 (AKT1) were screened out by PPI network analysis. A total of 166 pathways were obtained by enrichment of KEGG pathway, and 90 pathways closely related to the disease were screened out, including phosphatidylinositol-3-kinase-protein kinase B signaling pathway, mitogen-activated protein kinase signaling pathway, serotonin synapse, neurotrophic signaling pathway and Tolllike receptor signaling pathway. The results of molecular docking showed that the 10 main components in Erxiekang Emplastra had strong binding ability with 15 core target proteins and could form stable complexes. Conclusion All the transdermal components of Erxiekang Emplastra were obtained by HPLC-Q-TOF-MS/MS, and combined with network pharmacology and molecular docking technology, the potential material basis and mechanism of Erxiekang Emplastra in treating children's diarrhea with multicomponents, multi-targets and multi-channels were preliminarily determined.

R285.5

國家自然科學(xué)基金委員會(huì)重點(diǎn)項(xiàng)目（81830111）