Dantas Bruna Braga, Faheina-Martins Gláucia Veríssimo, Coulidiati Tangbadioa Hervé, Bomfim Caio César Barbosa, da Silva Dias Celidarque, Barbosa-Filho José Maria, Araújo Demetrius Antônio Machado
Department of Biotechnology, Center of Biotechnology, Federal University of Paraíba, Cidade Universitária, Campus I, 58051-900, João Pessoa, PB, Brazil.
J Nat Med. 2015 Apr;69(2):218-23. doi: 10.1007/s11418-014-0881-5. Epub 2015 Jan 24.
Curine is a natural alkaloid isolated from Chondrodendron platyphyllum and it has been reported that this alkaloid has vasodilatory and anti-inflammatory effects. The aim of this study is to analyze the cytotoxic effects of curine in cancer cell lines HL-60, K562, and HT-29, and in primary cultures of peripheral blood mononuclear cells (PBMC). Cells were treated with curine (from 3 to 15 µM) for 24 and 48 h. Cell viability was analyzed by the 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test and flow cytometry with propidium iodide (PI) assay. To assess the type of cell death induced in HL-60, the cell cycle, morphological, and biochemical alterations were analyzed, which were determined by differential staining with acridine orange/ethidium bromide, and annexin V/PI double-labeling and change in mitochondrial membrane potential assays. Curine demonstrated a potent cytotoxic effect on leukemic cell lines (HL-60 and K562). Its cytotoxic effects in HL-60 cells was related to plasma membrane damage and cell cycle arrest at the G1 phase from 43.4 ± 1.0 to 56.7 ± 1.4 % (p < 0.05). Curine (15 μM) also increased the apoptotic cells number by around 60 % in HL-60 cells and caused phosphatidylserine externalization, inducing about 57 % of apoptosis. Moreover, this alkaloid provoked 20 % of mitochondrial membrane depolarization. We conclude that curine presented a cytotoxic effect and induced apoptosis in HL-60 cells. Thus, it can be considered a promising pharmacological drug.
箭毒碱是从阔叶南美防己中分离出的一种天然生物碱，据报道，这种生物碱具有血管舒张和抗炎作用。本研究的目的是分析箭毒碱对癌细胞系HL-60、K562和HT-29以及外周血单个核细胞（PBMC）原代培养物的细胞毒性作用。细胞用箭毒碱（3至15μM）处理24小时和48小时。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）试验和碘化丙啶（PI）流式细胞术分析细胞活力。为了评估HL-60中诱导的细胞死亡类型，分析了细胞周期、形态和生化改变，这些改变通过吖啶橙/溴化乙锭鉴别染色、膜联蛋白V/PI双标记以及线粒体膜电位测定来确定。箭毒碱对白血病细胞系（HL-60和K562）表现出强大的细胞毒性作用。其在HL-60细胞中的细胞毒性作用与质膜损伤和细胞周期在G1期停滞有关，从43.4±1.0%至56.7±1.4%（p<0.05）。箭毒碱（15μM）还使HL-60细胞中的凋亡细胞数量增加约60%，并导致磷脂酰丝氨酸外化，诱导约57%的凋亡。此外，这种生物碱引起20%的线粒体膜去极化。我们得出结论，箭毒碱对HL-60细胞具有细胞毒性作用并诱导凋亡。因此，它可被认为是一种有前景的药理药物。
