目的 采用網(wǎng)絡(luò)藥理學(xué)方法預(yù)測(cè)黃瑞香發(fā)揮抗胃癌作用的潛在機(jī)制，并通過體外細(xì)胞實(shí)驗(yàn)進(jìn)行初步驗(yàn)證。方法 利用中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫和分析平臺(tái)（TCMSP）篩選黃瑞香的有效成分，通過Swiss Target Prediction數(shù)據(jù)庫和PharmMapper數(shù)據(jù)庫篩選獲得黃瑞香活性成分的相關(guān)靶點(diǎn)。通過GeneCards及DisGeNET數(shù)據(jù)庫檢索獲得胃癌疾病相關(guān)靶點(diǎn)。利用Cytoscape軟件構(gòu)建黃瑞香活性成分-靶點(diǎn)網(wǎng)絡(luò)；通過STRING、Metascape數(shù)據(jù)庫對(duì)成分和胃癌疾病交集靶點(diǎn)進(jìn)行蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)和基因本體（GO）注釋及京都基因與基因組百科全書（KEGG）通路富集分析。采用MTT實(shí)驗(yàn)考察黃瑞香異戊烯基黃酮成分構(gòu)樹黃酮醇F（broussoflavonol F）、daphnegiravone D、daphgiflavone C體外對(duì)胃癌MGC803細(xì)胞生長(zhǎng)的影響，采用實(shí)時(shí)熒光定量PCR方法檢測(cè)這3種異戊烯基黃酮類化合物作用48h后胃癌MGC803細(xì)胞表皮生長(zhǎng)因子受體（EGFR）、磷脂酰肌醇3-激酶（PI3K）和蛋白激酶B（Akt）基因的表達(dá)。結(jié)果 篩選得到黃瑞香70個(gè)活性成分，黃瑞香作用于胃癌的42個(gè)相關(guān)靶點(diǎn)，其中表皮生長(zhǎng)因子受體（EGFR）、鼠肉瘤病毒癌基因同源物B1（BRAF）、Ⅰ類磷脂酰肌醇-3激酶催化亞基α（PIK3CA）、酪氨酸激酶（MET）以及絲裂原活化蛋白激酶1（MAPK1）等為核心靶點(diǎn)。KEGG富集分析發(fā)現(xiàn)黃瑞香作用于胃癌主要富集在癌癥、PI3K/Akt以及絲裂原活化蛋白激酶（MAPK）等通路中?；钚詼y(cè)試結(jié)果發(fā)現(xiàn)，異戊烯基黃酮成分構(gòu)樹黃酮醇F、daphnegiravone D、daphgiflavone C對(duì)胃癌MGC803細(xì)胞生長(zhǎng)具有較好的抑制作用。同時(shí)，與對(duì)照組比較，構(gòu)樹黃酮醇F、daphnegiravone D、daphgiflavone C可顯著抑制胃癌MGC803細(xì)胞EGFR、PI3K和Akt基因的表達(dá)（P＜0.05、0.01、0.001）。結(jié)論 黃瑞香可能以異戊烯基黃酮為主要的抗胃癌活性成分，以EGFR、BRAF、PIKC3A、MET等蛋白為核心作用靶點(diǎn)，主要經(jīng)I3K/Akt以及MAPK等相關(guān)信號(hào)通路發(fā)揮治療胃癌的作用。

Objective To investigate the mechanism of Daphne giraldii in the treatment of gastric cancer by using the method of network pharmacology and cellular experiments. Methods Systematic Pharmacology Database and TCMSP database were used to obtain the chemical components and action targets of D.giraldii. Then, from the Swiss target prediction, PharmMapper, GeneCards and DisGeNET databases, the targets of chemical constituents from D.giraldii and gastric cancer related target information were searched. The active ingredient-target network of D.giraldii was further constructed by Cytoscape. Afterwards, STRING and Metascape databases were used for protein-protein interaction, GO and KEGG pathway enrichment analysis. MTT assay was used to detect the anti-tumor activity of broussoflavonol F, daphnegiravone D, daphgiflavone C. The expression of mRNA level of EGFR, PI3K and Akt of gastric cancer MGC803 cells were examined by using real-time fluorescence quantitative PCR (qRT-PCR) analysis after 48h treated by these three prenylated flavonoids. Results Totally 70 potential active ingredients of D.giraldii were obtained. And through screening, 42 targets of D.giraldii on gastric cancer were obtained. Among them, EGFR, BRAF, PIK3CA, MET and MAPK1 were the core targets, which were mainly enriched in cancer, PI3K/Akt and MAPK signaling pathways. MTT assay showed that three prenylated flavones (broussoflavonol F, daphnegiravone D, daphgiflavone C) had better activity against gastric cancer MGC803 cells growth, and they also significantly decreased the mRNA level of EGFR, PI3K and Akt. Conclusion The main antigastric cancer active component of D.giraldii may be prenylated flavonoids, with EGFR, BRAF, PIKC3A, MET and other proteins as core targets, and it mainly plays a role in the treatment of gastric cancer through PI3K/Akt and MAPK signaling pathways.

R285.5

國家級(jí)大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計(jì)劃支持項(xiàng)目（202110163030）；國家自然科學(xué)基金資助項(xiàng)目（81973528）