RESUMO
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.
Assuntos
Antineoplásicos/farmacologia , Membrana Celular/química , Membrana Celular/efeitos dos fármacos , Indigofera/química , Antineoplásicos/isolamento & purificação , Apoptose , Sobrevivência Celular/efeitos dos fármacos , China , Cromatografia , Biologia Computacional , Humanos , Células K562/química , Células K562/efeitos dos fármacos , Estruturas Vegetais/química , Proto-Oncogene MasRESUMO
Objective To screen potential active anti-cancer components of Brucea javanica.Methods This research has em-ployed comprehensive two dimensional chromatographic technology and cell membrane chromatographic technology simultaneously with mass spectrometry as detector .Results Adenosine and Bruceine B were found to be potentially anti-cancer active .Conclusion This study has combined the advantages of online , high speed and high throughput for the screen of potential active components of traditional Chinese medicine .