Mojarrab Mahdi, Emami Seyed Ahmad, Delazar Abbas, Tayarani-Najaran Zahra
Pharmaceutical Sciences Research Center, School of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Department of Pharmacognosy, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Iran J Pharm Res. 2017 Winter;16(1):221-229.
Considering multiple reports on cytotoxic activity of the genus and its phytochemicals, in the current study Lam. and the three components isolated from the plant were subjected to cytotoxic studies. Analytical fractionation of aerial parts for the first time was directed to the isolation of 7-hydroxy-8-(4-hydroxy-3-methylbutoxy) comarin (armenin), 8-hydroxy-7-(4-hydroxy-3-methylbutoxy) comarin (isoarmenin) and deoxylacarol. Cytotoxicity assessed with alamalBlue® assay and apoptosis was detected by PI staining and western blot analysis of Bax and PARP proteins. Extracts and all compounds exhibited cytotoxic activity against apoptosis-proficient HL-60 and apoptosis-resistant K562 cells, with the lowest cytotoxic activity on J774 cell line as non-malignant cell. Armenin as the most potent component decreased the viability of cell with IC50 of 22.5 and 71.1 µM for K562 and HL-60 cells respectively and selected for further mechanistic study. Armenin increased the sub-G1 peak in flow cytometry histogram of HL-60 and K562 treated cells and increase in the amount of Bax protein and the cleavage of PARP in comparison with the control after treatment for 48 h in K562 treated cells verified the apoptotic activity of the armenin. Taken together, according to the finding of this study armenin was introduced as a novel cytotoxic compound with apoptotic activity, which is encouraging for further mechanistic and clinical studies.
鉴于关于该属及其植物化学成分细胞毒性活性的多篇报道，在本研究中，对Lam.以及从该植物中分离出的三种成分进行了细胞毒性研究。首次对地上部分进行分析分离，旨在分离出7-羟基-8-(4-羟基-3-甲基丁氧基)香豆素（阿门菌素）、8-羟基-7-(4-羟基-3-甲基丁氧基)香豆素（异阿门菌素）和脱氧拉卡醇。用AlamalBlue® 检测法评估细胞毒性，通过PI染色以及对Bax和PARP蛋白进行蛋白质印迹分析来检测细胞凋亡。提取物和所有化合物对凋亡能力正常的HL-60细胞和凋亡抗性K562细胞均表现出细胞毒性活性，对作为非恶性细胞的J774细胞系的细胞毒性活性最低。阿门菌素作为最有效的成分，分别使K562和HL-60细胞的活力降低，IC50分别为22.5和71.1 µM，并被选用于进一步的机制研究。阿门菌素增加了HL-60和K562处理细胞的流式细胞术直方图中的亚G1峰，并且在K562处理细胞中处理48小时后，与对照相比，Bax蛋白量的增加和PARP的裂解证实了阿门菌素的凋亡活性。综上所述，根据本研究结果，阿门菌素被鉴定为一种具有凋亡活性的新型细胞毒性化合物，这为进一步的机制和临床研究提供了依据。
