Gajdus Jerzy, Kaczyński Zbigniew, Kawiak Anna, Lojkowska Ewa, Stefanowicz-Hajduk Justyna, Ochocka J Renata, Stepnowski Piotr
Department of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-308, Gdańsk, Poland.
Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Kładki 24, 80-822, Gdańsk, Poland.
Pharmacogn Mag. 2014 Apr;10(Suppl 2):S324-33. doi: 10.4103/0973-1296.133289.
Paris quadrifolia L. is a medicinal plant which contains steroidal saponins. The present study reports isolation and structural identification of six pennogenyl saponins obtained from P. quadrifolia rhizomes. The four spirostan saponins were obtained from P. quadrifolia for the first time. The cytotoxic effects of the sub-fractions and six compounds isolated from the plant extract were evaluated on tumour cells.
Ethanol extract from the rhizomes of P. quadrifolia were partinioned using column chromatography. The saponins were isolated from the obtained sub-fractions by isocratic RP HPLC and their structures were determined by means of 1D and 2D NMR spectroscopy and MALDI TOF MS. The cytotoxic effects of the sub-fractions and the isolated compounds were tested against human promyelocytic leukaemia cells (HL-60), human cervical adenocarcinoma cells (HeLa) and human breast cancer cells (MCF-7) using the [(3-(4,5-dimethylthiazol-2-yl)]-2,5-diphenyltetrazolium bromide (MTT) assay.
Six pennogenyl saponins were isolated from P. quadrifolia rhizomes: pennogenin 3-O-β-D-glucopyranoside (1), pennogenin 3-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranoside (2), pennogenin 3-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside (3), pennogenin 3-O-α-L-rhamnopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranoside (4), pennogenin 3-O-α-L-rhamnopyranosyl-(1→4)-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside (5), pennogenin 3-O-α-L-rhamnopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→4)-[α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside (6). Pennogenyl saponins 5 and 6 exhibited cytotoxic activity against HL-60, HeLa and MCF-7 tumour cells with IC50 values of 1.0 ± 0.04 μg/ml, 1.8 ± 0.072 μg/ml and 2.4 ± 0.096 μg/ml respectively, and 2.0 ± 0.08 μg/ml, 2.5 ± 0.125 μg/ml and 3.2 ± 0.128 μg/ml respectively.
Compounds 1-4 were isolated from this species for the first time.
四叶重楼是一种含有甾体皂苷的药用植物。本研究报道了从四叶重楼根茎中分离得到的六种原薯蓣皂苷元皂苷及其结构鉴定。四种螺旋甾烷皂苷首次从四叶重楼中获得。评估了植物提取物中分离得到的亚组分和六种化合物对肿瘤细胞的细胞毒性作用。
采用柱色谱法对四叶重楼根茎的乙醇提取物进行分离。通过等度反相高效液相色谱法从得到的亚组分中分离出皂苷，并通过一维和二维核磁共振光谱以及基质辅助激光解吸电离飞行时间质谱确定其结构。采用[3-(4,5-二甲基噻唑-2-基)]-2,5-二苯基四氮唑溴盐（MTT）法检测亚组分和分离得到的化合物对人早幼粒细胞白血病细胞（HL-60）、人宫颈腺癌细胞（HeLa）和人乳腺癌细胞（MCF-7）的细胞毒性作用。
从四叶重楼根茎中分离得到六种原薯蓣皂苷元皂苷：原薯蓣皂苷元3-O-β-D-吡喃葡萄糖苷（1）、原薯蓣皂苷元3-O-α-L-吡喃鼠李糖苷-(1→4)-β-D-吡喃葡萄糖苷（2）、原薯蓣皂苷元3-O-α-L-吡喃鼠李糖苷-(1→2)-β-D-吡喃葡萄糖苷（3）、原薯蓣皂苷元3-O-α-L-吡喃鼠李糖苷-(1→4)-α-L-吡喃鼠李糖苷-(1→4)-β-D-吡喃葡萄糖苷（4）、原薯蓣皂苷元3-O-α-L-吡喃鼠李糖苷-(1→4)-[α-L-吡喃鼠李糖苷-(1→2)]-β-D-吡喃葡萄糖苷（5）、原薯蓣皂苷元3-O-α-L-吡喃鼠李糖苷-(1→4)-α-L-吡喃鼠李糖苷-(1→4)-[α-L-吡喃鼠李糖苷-(1→2)]-β-D-吡喃葡萄糖苷（6）。原薯蓣皂苷元皂苷5和6对HL-60、HeLa和MCF-7肿瘤细胞具有细胞毒性活性，IC50值分别为1.0±0.04μg/ml、1.8±0.072μg/ml和2.4±0.096μg/ml，以及2.0±0.08μg/ml、2.5±0.125μg/ml和3.2±0.128μg/ml。
化合物1-4首次从该物种中分离得到。
