Lc-ms And Drug Screening In Tcm Toxicity Studies In Application Exploration

With the increasingly widespread use of Traditional Chinese medicine (TCM), the Adverse reactions of TCM are gradually emerging, which not only makes the toxicology studies of TCM become more and more important, but also urgently require modernization of TCM toxicity studies and the establishment of standardized research platforms. To aim directly at eighteen incompatible medicaments, we carried out the two levels of exploratory study of TCM toxicity with LC-MS technology and drug screening technology, which investigate the changing of TCM toxic components of the chemical compatibility substance and drug metabolizing enzymes on the metabolism of toxic components of Chinese medicine. The objective of this research was to provide experimental evidences for the mode of drug interaction based on CYP and the mechanism of incomepatible medicaments, as well as constructing TCM toxicology research platform.The material basis of TCM clinical efficacy or toxicity is the sum of its chemical composition. Chemical composition of TCM is very complex, so it is difficult to completely analyse the chemical composition spectrum and toxicity component spectrum of opposing herbal compatibility by conventional methods. Howere, UPLC/Q-TOF-MS technology overcomed these difficulties. Our results indicate pseudo-Jervine, verazine and verazine elevated significantly, but angeloyl zygadenine, veratramine alkali and germidine decreased after co-decocting Veratrum nigrum L and Radix Glehniae. After co-decocting Veratrum nigrum L and Radix Codonopsis, only pseudo-Jervine content was increased, other alkaloids content of Veratrum nigrum L was down regulated. The above results indicated that more poisonous and less effective action existed in compatibility of Veratrum nigrum L and Radix Glehniae, however, this phenomenon was not obvious between Veratrum nigrum L and Radix Codonopsis. This study preliminary explained the chemical basis of increasing toxicity after compatibility.TCM drug interaction is very common because of combination therapy. Most of the mechanism of the TCM drug interaction is related to drug metabolism, especially related to cytochrome P450 enzymes. So, it is a useful idea and method, which development a CYP-based drug screening technology for understanding the TCM toxicology. This study was to construct a secreted luciferase reporter gene, with two-distal enhancer and proximal promoter tandemly ligated. A stably transfected HepG2 cells, which can predict the compounds'ability of CYP3A4 induction or inhibition by direct supernatant detection, will overcome the shortage of pGL series dual luciferase reporter gene. The secreted luciferase reporter gene with a single distal enhancer, had the higher response to RIF than a single distal enhancer pGL4.17 vector. Compairing with a single distal enhancer secreted luciferase reporter gene, double distal enhancer secreted luciferase reporter gene had not only the higher absolute fluorescence values to be significantly than that secreted remote enhancer luciferase reporter gene, but also further improved performance on RIF induction. Stable cell line transfected with new reporter gene is accurate and reliable, which is verified by positive inducers of CYP3A4 and inhibiting drugs of PXR.Using this in vitro screening system based on PXR-CYP3A4 pathway,70 Chinese herb compounds were screened by this method, and some compounds with potential capacity of CYP3A4 induction and inhibition were obtained. This technology platform provides new ideas and methods for research of TCM toxicity based on in vivo drug interactions.