目的 研究土当归Angelica gigas的化学成分，对分离得到的单体化合物进行抗前列腺癌活性评价。方法 利用硅胶柱色谱、半制备高效液相色谱等方法对土当归醋酸乙酯萃取物进行系统分离纯化，综合运用核磁共振、高分辨质谱等多种波谱手段对得到的单体化合物进行结构鉴定。采用CCK-8法测定化合物抗前列腺癌活性，利用网络药理学预测土当归中活性成分抗前列腺癌的靶点、功能及通路，通过蛋白质印迹实验进行验证。结果 从土当归醋酸乙酯部位分离得到33个化合物，依次鉴定为伞形花内酯（1）、茵芋苷（2）、东莨菪素（3）、7-去甲基软木花椒素（4）、6,7-二甲氧基香豆素（5）、devenyol（6）、佛手柑内酯（7）、异欧前胡素（8）、蛇床素（9）、欧前胡素（10）、花椒毒酚（11）、花椒毒素（12）、紫花前胡苷元（13）、异紫花前胡内酯（14）、紫花前胡苷（15）、异补骨脂素（16）、噢洛内酯（17）、(±)-vaginidiol（18）、二氢山芹醇当归酸酯（19）、花椒内酯（20）、紫花前胡醇（21）、紫花前胡素（22）、紫花前胡醇当归酯（23）、(＋)-lomatin（24）、(＋)-顺式凯林内酯（25）、儿茶素（26）、槲皮素（27）、杨梅素（28）、烟酸（29）、阿魏酸（30）、绿原酸（31）、5-羟甲基糠醛（32）、5,5¢-二丁氧基-2,2¢-双环呋喃（33）。化合物1、4、6、8、12、13、19、21～24表现出抑制前列腺癌细胞增殖作用，进一步对抑癌作用最显著的5个化合物（1、4、21～23）进行抗前列腺癌活性评价，其中化合物22、23抑制前列腺肿瘤作用最强。通过网络药理学预测获得的土当归中主要活性成分（化合物1、4、21～23）抗前列腺癌核心靶点11个，包括蛋白激酶B（threonine kinase，AKT）、表皮生长因子受体（epidermal growth factor receptor，EGFR）、细胞周期蛋白D1（cyclin D1，CCND1）、前列腺素内过氧化物合成酶2（prostaglandin-endoperoxide synthase 2，PTGS2）等；KEGG通路富集筛选得到信号通路涉及癌症通路、磷脂酰肌醇3-激酶/蛋白激酶B（phosphatidylinositol 3-kinase/protein kinase B，PI3K/AKT）通路、EGFR信号通路等。经Western blotting验证土当归中活性成分通过抑制PI3K/AKT通路中关键蛋白AKT的表达发挥抗前列腺癌的作用。结论 化合物6、16、17、24、33为首次从该属植物中发现，化合物18、25、32为首次从该种植物中发现。化合物1、4、21～23表现出较好的抑制前列腺癌细胞增殖作用，其中化合物22和23的抑制作用最强，其通过抑制PI3K/AKT通路中关键蛋白AKT的表达从而发挥抗前列腺癌的作用。

Objective To study the chemical constituents of of Angelica gigas, and evaluate the activity of the isolated monomer compounds against prostate cancer. Methods The ethyl acetate extract of A. gigas was systematically isolated and purified using silica gel column chromatography and semi-preparative HPLC. The structures of the isolated monomer compounds were characterized by a combination of spectroscopic methods, including nuclear magnetic resonance, high resolution mass spectrometry. The anti-prostate cancer activity of the compounds was evaluated using the CCK-8 assay, and network pharmacology was employed to predict the targets, functions, and pathways of the active ingredients of A. gigas against prostate cancer, which was further verified by Western blotting (WB) experiments. Results A total of 33 compounds were isolated from the ethyl acetate fraction of A. gigas and identified as umbelliferone (1), skimmin (2), scopoletin (3), 7-demethylsuberosine (4), 6,7-dimethoxycoumarin (5) devenyol (6), bergapten (7), isoimperatorin (8), cnidicin (9), imperatorin (10), xanthotoxol (11), xanthotoxin (12), nodakenetin (13), marmesin (14), nodakenin (15), isopsoralen (16), oroselol (17), (±)-vaginidiol (18), columbianadin (19), xanthyletin (20), decursinol (21), decursin (22), decursinol angelate (23), (＋)-lomatin (24) (＋)-cis-khellactone (25), catechin (26), quercetin (27), myricetin (28), nicotinic acid (29), ferulic acid (30), chlorogenic acid (31), 5-(hydroxymethyl)furfural (32), 5,5¢-dibutoxy-2,2¢-bifuran (33). Compounds 1, 4, 6, 8, 12, 13, 19, and 21—24 exhibited inhibition of prostate cancer cell proliferation. The five compounds with the most significant cancer inhibition effects (1, 4, 21—23) were further evaluated for their anti-prostate cancer activities at multiple concentrations. Among them, compounds 22 and 23 showed the strongest inhibition of prostate tumors. A total of 11 core anti-prostate cancer targets of the chemical components in A. gigas were identified through network pharmacology prediction, including threonine kinase (AKT), epidermal growth factor receptor (EGFR), cyclin D1 (CCND1), prostaglandin-endoperoxide synthase 2 (PTGS2), and others. KEGG pathway enrichment screening revealed signaling pathways involved in the cancer pathway, the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway, and the EGFR signaling pathway. Western blotting analysis confirmed that the active ingredients in A. gigas exerted anti-prostate cancer effects by inhibiting the expression of AKT, a key protein in the PI3K/AKT pathway. Conclusion Compounds 6, 16, 17, 24, and 33 were identified for the first time from this genus, while compounds 18, 25, and 32 were found for the first time from this species. Compounds 1, 4, 21—23 showed better inhibition of prostate cancer cell proliferation, with compounds 22 and 23 having the strongest inhibitory effect, which demonstrated their anti-prostate cancer effects by inhibiting the expression of AKT, a key protein in the PI3K/AKT pathway.

R284.1

广东省自然科学基金项目（2021A1515011485）；辽宁省中医药多学科交叉创新团队项目（LNZYYCXTD-JCCX-002）