Characterization of anti-leukemia components from Indigo naturalis using comprehensive two-dimensional K562/cell membrane chromatography and in silico target identification.
Characterization of anti-leukemia components from Indigo naturalis using comprehensive two-dimensional K562/cell membrane chromatography and in silico target identification.
复制标题
使用全面的二维 K562/细胞膜色谱法和计算机靶点识别来表征青黛的抗白血病成分
DOI：
10.1038/srep25491
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发表时间：
2016-05-06
影响因子：
4.6
通讯作者：
Zhang L
中科院分区：
文献类型：
--
作者：
Wu X;Chen X;Dan J;Cao Y;Gao S;Guo Z;Zerbe P;Chai Y;Diao Y;Zhang L
中医学经过几千年的发展，在大量临床实践的基础上形成了完整的理论体系。然而，中药中的主要成分尚未完全确定，其确切的作用机制和靶点尚未阐明。本研究采用K562/细胞膜全二维色谱系统和计算机靶点识别相结合的新方法，对中药靛蓝中的活性成分及其靶点进行了研究。三个活性成分，靛玉红，色胺酮和异鼠李素，成功地表征和他们的抗白血病细胞活力和细胞凋亡实验验证。选择具有未定义的癌症相关靶标的异鼠李素用于计算机模拟靶标鉴定。原癌基因酪氨酸蛋白激酶（Src）是其膜靶蛋白，Src与异鼠李素的解离常数（Kd）为3.81 μM。异鼠李素的抗白血病作用可能是通过Src诱导G2/M期细胞阻滞而实现的。结果表明，该方法能够有效地表征中药活性成分及其作用靶点，为深入研究中药的复杂作用机制提供了新的思路。
Traditional Chinese Medicine (TCM) has been developed for thousands of years and has formed an integrated theoretical system based on a large amount of clinical practice. However, essential ingredients in TCM herbs have not been fully identified, and their precise mechanisms and targets are not elucidated. In this study, a new strategy combining comprehensive two-dimensional K562/cell membrane chromatographic system and in silico target identification was established to characterize active components from Indigo naturalis, a famous TCM herb that has been widely used for the treatment of leukemia in China, and their targets. Three active components, indirubin, tryptanthrin and isorhamnetin, were successfully characterized and their anti-leukemia effects were validated by cell viability and cell apoptosis assays. Isorhamnetin, with undefined cancer related targets, was selected for in silico target identification. Proto-oncogene tyrosine-protein kinase (Src) was identified as its membrane target and the dissociation constant (Kd) between Src and isorhamnetin was 3.81 μM. Furthermore, anti-leukemia effects of isorhamnetin were mediated by Src through inducing G2/M cell cycle arrest. The results demonstrated that the integrated strategy could efficiently characterize active components in TCM and their targets, which may bring a new light for a better understanding of the complex mechanism of herbal medicines.