目的 利用網(wǎng)絡(luò)藥理學(xué)方法和分子對(duì)接技術(shù)探索四逆散治療毒性彌漫性甲狀腺腫（Graves）病的潛在活性成分、作用靶點(diǎn)和代謝通路，為四逆散治療Graves病的臨床應(yīng)用和深入闡明四逆散干預(yù)Graves病的作用機(jī)制提供理論參考。方法 運(yùn)用TCMSP平臺(tái)及查閱文獻(xiàn)篩選四逆散的主要活性成分及作用靶點(diǎn)。以“Graves’ disease”為關(guān)鍵詞，挖掘GeneCards、CTD、DisGeNET、OMIM、TTD和DrugBank數(shù)據(jù)庫(kù)中Graves病的相關(guān)靶點(diǎn)。利用Cytoscape軟件構(gòu)建“中藥–成分–靶點(diǎn)–疾病”網(wǎng)絡(luò)，運(yùn)用String平臺(tái)構(gòu)建蛋白互相作用（PPI）網(wǎng)絡(luò)。運(yùn)用Metascape平臺(tái)對(duì)共同靶點(diǎn)通過(guò)GO數(shù)據(jù)庫(kù)和KEGG數(shù)據(jù)庫(kù)進(jìn)行注釋和富集分析。在RCSB PDB和PubChem數(shù)據(jù)庫(kù)中獲得成分和靶點(diǎn)的結(jié)構(gòu)，利用pymol軟件和autodock vina軟件進(jìn)行分子對(duì)接。結(jié)果 四逆散治療Graves病的主要活性成分可能為槲皮素、山柰酚、芹菜素、異鼠李素、柚皮素、異甘草素、木犀草素、葛根素等，PPI網(wǎng)絡(luò)中關(guān)鍵靶點(diǎn)為AKT1、TNF、INS、IL6、VEGFA、TP53、JUN、CASP3、IL1B、MAPK3等。GO與KEGG分析主要代謝通路可能有PI3K-Akt信號(hào)通路、TNF信號(hào)通路、IL-17信號(hào)通路、HIF-1信號(hào)通路、p53信號(hào)通路、細(xì)胞凋亡、Th17細(xì)胞分化、Toll樣受體信號(hào)通路、甲狀腺激素信號(hào)通路、VEGF信號(hào)通路等。四逆散治療Graves病的作用機(jī)制可能有調(diào)節(jié)免疫失衡、抑制細(xì)胞增殖和促進(jìn)凋亡、抗氧化應(yīng)激、抑制甲狀腺血管新生和減輕外周組織器官損傷等。分子對(duì)接結(jié)果顯示四逆散主要成分與治療Graves病的關(guān)鍵靶點(diǎn)結(jié)合活性較高且對(duì)接構(gòu)象較穩(wěn)定。結(jié)論 四逆散可能通過(guò)多成分、多靶點(diǎn)影響多條代謝通路發(fā)揮治療Graves病的作用，為四逆散治療Graves病的作用機(jī)制實(shí)驗(yàn)研究提供參考。

Objective The potential active components, action targets and metabolic pathways of Sinisan in treatment of Graves' disease were explored by using network pharmacology and molecular docking technology, so as to provide theoretical reference for the clinical application of Sinisan in treatment of Graves' disease and further elucidation of the mechanism of action of Sinisan in the intervention of Graves' disease. Methods The main active components and targets of Sinisan were screened by TCMSP platform and literature review. With “Graves' disease” as the key word, the related targets of Graves' disease were extracted from GeneCards, CTD, DisGeNET, OMIM, TTD and DrugBank databases. Cytoscape software was used to construct the network of “TCM - component - target - disease”, and String platform was used to construct PPI network. Metascape platform was used to annotate and enrich common targets through GO database and KEGG database. The structures of components and targets were obtained from RCSB PDB and PubChem databases, and molecular docking was performed using Pymol software and Autodock Vina software. Results The main active ingredients of Sinisan in treatment of Graves' disease may be quercetin, kaempferol, apigenin, isornetin, naringin, isorritin, luteolin, puerarin, etc. The key targets in PPI network are AKT1, TNF, INS, IL-6, VEGFA, TP53, JUN, CASP3, IL-1B, MAPK3, etc. The main metabolic pathways analyzed by GO and KEGG may include PI3K-Akt signaling pathway, TNF signaling pathway, IL-17 signaling pathway, HIF-1 signaling pathway, p53 signaling pathway, apoptosis, Th17 cell differentiation, Toll-like receptor signaling pathway, thyroid hormone signaling pathway, VEGF signaling pathway, etc. The mechanism of Sinisan in treating Graves' disease may include regulating immune imbalance, inhibiting cell proliferation and promoting apoptosis, resisting oxidative stress, inhibiting thyroid angiogenesis and alleviating peripheral tissue and organ injury. The molecular docking results showed that the main active components of Sinisan had high binding activity and stable docking conformation with the key target of Graves' disease. Conclusion Sinisan may affect multiple metabolic pathways through multiple components and multiple targets, which can provide reference for experimental study on the mechanism of action of Sinisan in the treatment of Graves' disease.

R977

國(guó)家自然科學(xué)基金青年項(xiàng)目(82004337);國(guó)家自然科學(xué)基金面上項(xiàng)目(81774293);北京中醫(yī)藥大學(xué)新教師啟動(dòng)基金項(xiàng)目(2020-JYB-XJSJJ-002)