目的 通過網(wǎng)絡(luò)藥理學(xué)和實(shí)驗(yàn)驗(yàn)證方法探討雷公藤治療黑色素瘤的活性成分、作用靶點(diǎn)及信號通路，分析雷公藤對黑色素瘤的作用機(jī)制。方法 通過TCMSP數(shù)據(jù)庫檢索雷公藤的有效成分及靶點(diǎn)，借助GeneCards、OMIM數(shù)據(jù)庫獲取中藥與疾病的交集靶點(diǎn)。采用Cytoscape 3.7.1進(jìn)行拓?fù)浞治?，?gòu)建“藥物活性成分–疾病靶點(diǎn)”網(wǎng)絡(luò)圖。運(yùn)用String平臺構(gòu)建共同靶點(diǎn)蛋白相互作用（PPI）網(wǎng)絡(luò)和條形圖，篩選出核心靶點(diǎn)。進(jìn)行靶點(diǎn)基因富集分析，完成核心靶標(biāo)與成分的分子對接。利用CCK-8及qPCR方法驗(yàn)證核心單體成分對黑色素瘤及治療靶點(diǎn)的作用。結(jié)果 雷公藤有效成分51個，其中最優(yōu)成分是雷公藤甲素、山柰酚。雷公藤甲素對黑色素瘤細(xì)胞具有明顯抑制作用，且顯著降低核心治療靶點(diǎn)蛋白激酶B1（Akt1）、血管內(nèi)皮生長因子A（VEGFA）的mRNA表達(dá)（P＜0.01、0.001）。結(jié)論 雷公藤治療黑色素瘤具有多靶點(diǎn)、多通路的特點(diǎn)，其中雷公藤甲素是其核心藥用成分，該研究為雷公藤抗黑色素瘤提供了實(shí)驗(yàn)依據(jù)。

Objective To predict the active ingredients, targets and signaling pathways of Tripterygium wilfordii in treatment of melanoma by network pharmacology and experimentation, and to analyze the mechanism of Tripterygium wilfordii in treatment of melanoma. Methods To search the active components and targets of Tripterygium wilfordii from TCMSP database, and the intersection targets of traditional Chinese medicine and diseases were obtained from GeneCards and OMIM databases. Topological analysis was carried out on Cytoscape 3.7.1 to construct the “drug active ingredients - disease targets” network diagram. The interaction network and bar graph of common target protein were constructed with String platform, and the core target was screened out. Target gene enrichment analysis was carried out to complete the molecular docking of core targets and components. CCK8 and qPCR methods were used to verify the effects of core monomer components on melanoma and therapeutic targets. Results There were 51 active components of Tripterygium wilfordii. The best ingredient are kaempferol and triptolide. It has a significant inhibitory effect on melanoma cells, and significantly reduces the mRNA expression of core therapeutic targets Akt1 and VEGFA. Conclusion Tripterygium wilfordii has the characteristics of multi-target and multi-pathway in treatment of melanoma, and triptolide is its core medicinal component. This study provides experimental basis for Tripterygium wilfordii anti-melanoma.

R979.1

河南省科技研發(fā)計(jì)劃聯(lián)合基金（優(yōu)勢學(xué)科培育類）項(xiàng)目（222301420060）；河南省自然科學(xué)基金-優(yōu)秀青年科學(xué)基金項(xiàng)目（212300410057）