College of Pharmaceutical Sciences, Southwest University, 2 Tiansheng Road, Beibei District, Chongqing 400716, China.
College of Pharmaceutical Sciences, Southwest University, 2 Tiansheng Road, Beibei District, Chongqing 400716, China.
Phytomedicine. 2020 Mar;68:153142. doi: 10.1016/j.phymed.2019.153142. Epub 2019 Dec 4.
The dried heartwood of Caesalpinia sappan L. is traditionally prescribed in the formula of traditional Chinese medicine (TCM) for the treatment of acute myeloid leukemia (AML), while nothing is yet known of the active fractions and the underlying mechanisms.
This study aims to investigate the effect of the ethyl acetate extract of the dried heartwood of Caesalpinia sappan L. (C-A-E) on induction of apoptosis and promotion of differentiation in vitro and anti-AML activity in vivo.
STUDY DESIGN/METHODS: The aqueous extract was sequentially separated with solvents of increasing polarity and the active fraction was determined through the inhibition potency. The inhibition of the active fraction on cell viability, proliferation and colony formation was performed in different AML cells. Induction of apoptosis and the promotion of differentiation were further determined. Then, the level of the reactive oxygen species (ROS) and its potential role were assessed. Finally, anti-AML activity was evaluated in NOD/SCID mice.
C-A-E exhibited the highest inhibition on the cell viability of HL-60 cells. Meanwhile, C-A-E significantly suppressed the proliferation and the colony formation ability of HL-60 and Kasumi-1 cells. Moreover, C-A-E significantly induced the apoptosis, which was partially reversed by Z-VAD-FMK. C-A-E also reduced the level of mitochondrial membrane potential, promoted the release of cytochrome C, decreased the Bcl-2/Bax ratio, and promoted the cleavage of caspase-9 and -3. In addition, Mdivi-1 (mitochondrial fission blocker) remarkably reduced the apoptosis caused by C-A-E. Meanwhile, C-A-E also induced the expression of Mff and Fis1 and increased the location of Drp1 in mitochondria. Furthermore, C-A-E obviously promoted the differentiation of AML cells characterized by the typic morphological changes, the increased NBT positive cells, as well as the increased CD11b and CD14 levels. Notably, C-A-E significantly enhanced the intracellular ROS level. Moreimportantly, C-A-E-mediated apoptosis and differentiation of HL-60 cells was significantly mitigated by NAC. Additionally, C-A-E also exhibited an obvious anti-AML effect in NOD/SCID mice with the injection of HL-60 cells.
C-A-E exhibited an inhibitory effect on AML cells by inducing mitochondrial apoptosis and promoting differentiation, both of which were highly correlated to the activation of ROS.
苏木的干燥心材在传统中药（TCM）配方中被用于治疗急性髓系白血病（AML），但其有效部位和作用机制尚不清楚。
本研究旨在探讨苏木乙酸乙酯提取物（C-A-E）在体外诱导细胞凋亡和促进分化以及体内抗 AML 活性方面的作用。
研究设计/方法：将水提物依次用极性递增的溶剂分离，通过抑制活力确定活性部位。在不同的 AML 细胞中，测定活性部位对细胞活力、增殖和集落形成的抑制作用。进一步测定诱导细胞凋亡和促进分化的作用。然后，评估活性氧（ROS）的水平及其潜在作用。最后，在 NOD/SCID 小鼠中评估抗 AML 活性。
C-A-E 对 HL-60 细胞的细胞活力抑制作用最强。同时，C-A-E 显著抑制 HL-60 和 Kasumi-1 细胞的增殖和集落形成能力。此外，C-A-E 显著诱导细胞凋亡，Z-VAD-FMK 部分逆转了这一作用。C-A-E 还降低了线粒体膜电位水平，促进了细胞色素 C 的释放，降低了 Bcl-2/Bax 比值，并促进了 caspase-9 和 -3 的裂解。此外，Mdivi-1（线粒体分裂阻断剂）显著减少了 C-A-E 引起的细胞凋亡。同时，C-A-E 还诱导了 Mff 和 Fis1 的表达，并增加了 Drp1 在线粒体中的定位。此外，C-A-E 明显促进了 AML 细胞的分化，表现为典型的形态变化、增加的 NBT 阳性细胞以及 CD11b 和 CD14 水平的增加。值得注意的是，C-A-E 明显增加了细胞内 ROS 水平。更重要的是，NAC 显著减轻了 C-A-E 介导的 HL-60 细胞凋亡和分化。此外，C-A-E 对注射 HL-60 细胞的 NOD/SCID 小鼠也表现出明显的抗 AML 作用。
C-A-E 通过诱导线粒体凋亡和促进分化对 AML 细胞发挥抑制作用，这两种作用都与 ROS 的激活密切相关。
