钩吻-玉叶金花调控核受体PXR/CAR表达“相畏”减毒的分子机制研究

课题组前期完成的国家自然科学基金表明：钩吻配玉叶金花能减毒存效，与转运体P-gp外排、代谢酶CYP2E1代谢吲哚类生物碱有关。现已知核受体PXR/CAR负责外源性毒物或内源毒性脂类的解毒和排泌过程，能有效调节ABC转运体及CYP450代谢酶。因而研究提出“钩吻畏玉叶金花是通过调控核受体PXR/CAR来上调药物代谢酶及转运体的活性，而达到有效减缓钩吻毒性的目的”假说，对相畏是否通过核受体PXR/CAR介导的CYP450酶-ABC转运体途径展开探讨，通过正常与PXR/CAR转染细胞、中毒与解毒C57BL/6小鼠，检测其上游核受体（PXR、CAR）、下游CYP450酶（CYP2E1、CYP1A2、CYP2B6、CYP2C9、CYP3A4）与ABC转运体（P-gp、MRP2）基因与蛋白表达，从不同机能状态层面诠释相畏的作用靶点及分子机制。不仅阐明了相畏减毒的特色和优势，又为相畏减毒研究提供新思路。

The experimental data from the National Natural Science Foundation of China have already shown that the detoxification and analgesic efficacy of Gelsemium elegans combined with Mussaenda pubescens were closely related to efflux of indole alkaloids by P-gp and metabolism of indole alkaloids by CYP2E1. It is generally acknowledged that nuclear receptor PXR/CAR is in charge of detoxification and excretion process of exogenous toxins or endogenous lipid. They can effectively adjust CYP450 metabolic enzymes and ABC transporter. Therefore this research hypothesis is put forward: the detoxification mechanism of mutual restraint between Gelsemium elegans and Mussaenda pubescens works by regulating of CYP450 enzyme-ABC transporter through PXR/CAR nuclear receptor. The research is carried out to check that whether the detoxification mechanism is connected with CYP450 enzyme-ABC transporter regulated by PXR/CAR nuclear receptor. The therapeutic targets and detoxification mechanism will be elucidated from different body state between normal cells and PXR/CAR transfected cells, toxication mice and detoxification mice by determinating gene and protein expression levels of upstream nuclear receptors (PXR, CAR), downstream CYP450 enzymes (CYP2E1, CYP1A2, CYP2B6, CYP2C9, CYP3A4) and ABC transporter (P-gp, MRP2). Thus, the study will clarify the characteristics and advantages of detoxification research by mutual restraint, and provide a new method for the detoxification research of mutual restraint in traditional Chinese medicine.