PDF(Pubmed)
Abstract：
Ammi majus L. (Apiaceae) is a medicinal plant with a well-documented history in phytotherapy. The aim of the present work was to isolate isopimpinellin (5,8-methoxypsoralen; IsoP) from the fruit of this plant and evaluate its biological activity against selected tumor cell lines. The methanol extract obtained with the use of an accelerated solvent extraction (ASE) method was the most suitable for the quantitative analysis of coumarins in the A. majus fruit matrix. The coumarin content was estimated by RP-HPLC/DAD, and the amount of IsoP was found to be 404.14 mg/100 g dry wt., constituting 24.56% of the total coumarin fraction (1.65 g/100 g). This, along with the presence of xanthotoxin (368.04 mg/100 g, 22.36%) and bergapten (253.05 mg/100 g, 15.38%), confirmed A. majus fruits as an excellent source of these compounds. IsoP was isolated (99.8% purity) by combined liquid chromatography/centrifugal partition chromatography (LC/CPC) and tested for the first time on its antiproliferative activity against human colorectal adenocarcinoma (HT29, SW620), osteosarcoma (Saos-2, HOS), and multiple myeloma (RPMI8226, U266) cell lines. MTT assay results (96 h incubation) demonstrated a dose- and cell line-dependent decrease in cell proliferation/viability, with the strongest effect of IsoP against the Saos-2 cell line (IC50; 42.59 µM), medium effect against U266, HT-29, and RPMI8226 (IC50 = 84.14, 95.53, and 105.0 µM, respectively), and very weak activity against invasive HOS (IC50; 321.6 µM) and SW620 (IC50; 711.30 µM) cells, as well as normal human skin fibroblasts (HSFs), with IC50; 410.7 µM. The mechanistic study on the Saos-2 cell line showed that IsoP was able to reduce DNA synthesis and trigger apoptosis via caspase-3 activation. In general, IsoP was found to have more potency towards cancerous cells (except for HOS and SW620) than against healthy cells. The Selective Index (SI) was determined, underlining the higher selectivity of IsoP towards cancer cells compared to healthy cells (SI = 9.62 against Saos-2). All these results suggest that IsoP might be a promising molecule in the chemo-prevention and treatment of primary osteosarcoma.
摘要：
AmmimajusL.（Apiaceae）是一种药用植物，在植物疗法方面有着悠久的历史。本工作的目的是从该植物的果实中分离异impinellin（5,8-甲氧基补骨脂素；IsoP），并评估其针对选定肿瘤细胞系的生物学活性。使用加速溶剂萃取（ASE）方法获得的甲醇提取物最适合用于A.majus果实基质中香豆素的定量分析。通过RP-HPLC/DAD估算香豆素含量，发现IsoP的量为404.14mg/100g干重。,占香豆素总分数的24.56%(1.65克/100克)。这个,随着黄曲霉毒素的存在（368.04毫克/100克，22.36%)和bergapten(253.05毫克/100克，15.38%），确认A.majus水果是这些化合物的极好来源。通过联合液相色谱/离心分配色谱（LC/CPC）分离出IsoP（纯度为99.8％），并首次测试了其对人结肠直肠腺癌的抗增殖活性（HT29，SW620），骨肉瘤（Saos-2，HOS），和多发性骨髓瘤（RPMI8226，U266）细胞系。MTT分析结果（96小时孵育）表明细胞增殖/活力的剂量和细胞系依赖性降低，IsoP对Saos-2细胞系的作用最强（IC50;42.59µM），对U266、HT-29和RPMI8226的中等效果(IC50=84.14、95.53和105.0µM，分别)，对侵袭性HOS（IC50;321.6µM）和SW620（IC50;711.30µM）细胞的活性非常弱，以及正常人皮肤成纤维细胞（HSF），IC50；410.7µM。对Saos-2细胞系的机制研究表明，IsoP能够减少DNA合成并通过caspase-3激活引发细胞凋亡。总的来说,发现IsoP对癌细胞(HOS和SW620除外)比对健康细胞具有更大的效力。确定了选择性指数(SI)，强调与健康细胞相比，IsoP对癌细胞的选择性更高（SI=9.62对Saos-2）。所有这些结果表明，IsoP可能是原发性骨肉瘤的化学预防和治疗中的有前途的分子。
