High Throughput Screening Method Of Identifying PXR Receptor Agonist From Chinese Herbs

Objective: In this study, we screened the monomer components. of traditional Chinese medicinesby useing PXR-CYP3A stably transfected HepG2cell strain which was completed by our laboratory.First the stability of the cell lines and cell culture conditions were detected,Then some Chinesemedicines’ water decoction and monomer components were detected on the cell lines to study theability of PXR induction, to study the opposite effect mechanism of “various parameters Xin Shaorebel Veratryl ".Then118kinds of Chinese medicine monomers were screeninged, check theinduced and inhibition of the ingredients. Finally, we oberseved the hepatoprotective effect byDeoxyschizandrin in excessive LCA induced liver injury mice.Method: Stably transfected cell lines culture methods,secreted Gaussia luciferase kit and Victor Xmicroplate reader were used incomponents screening examination. Finally, we oberseved thehepato-protective effect by Deoxyschizandrin in excessive LCA induced liver injury mice.Results: We found processing compound activated carbon adsorption FBS.,processing time of48hours and RIF dosing concentration of5μM.At the same time,we found: reaction kit in the strongacid-base conditions, Gaussia luciferase remained stable, but cell growth was inhibited, in ahigh-DMSO conditions, the secretion of the cells and the luciferase remained stable. Inspectionfound that situation by induction on the various parameters Xin Shao rebel quinoa Lo TCMdecoction and monomer composition: Ginseng in the12.5mg/ml, North Shashen and Adenophoraeat a concentration of50mg/ml, Asarum in the concentration was10mg/ml, the strongest inductionof CYP3A4expression. Veratryl and white peony root of the strongest induction in multiples ofconcentration for both the initial concentration. By observing the cell growth of the kind of hole inthe high concentration of traditional Chinese medicine plus, Veratryl group slow cell growth, celldeath happens. Ginseng, Codonopsis, Salvia and other main component of the standard screening and found that the main ingredients of the ginseng-induced multiples are lower than the rifampingroup. Panaxatriol ginsenoside the Rf20(S)-Ginsenoside Rg2potential inhibition of CYP3A skills,20(S)-original panaxatriol,20(S)-ginsenoside F2induced multiples higher than the DMSOgroup. Sesamin, chrysophanol, ursolic acid, sodium Danshensu, protocatechuic acid, tanshinone ⅡA sulfonate also have different levels of induction and suppression. Investigation of traditionalChinese medicine monomer tetramethylpyrazine, one hundred autumn Li alcohol, and othercompounds activate PXR in varying degrees, with potential for enzyme induction effect;compounds baicalin, ginsenoside Rg1, oxymatrine, ginseng saponin Rb compounds in10μM underthe conditions of varying degrees of inhibition of PXR activity. Overall the results of animalexperiments, Deoxyschizandrin each dose group can effectively reverse the LCA-induced liverinjury in mouse peripheral blood ALT and AST increased, histological examination revealed thatmice given Deoxyschizandrin hepatocytes of the degree of necrosis have markedly improved, andDeoxyschizandrin expression is related to the synthesis and metabolism in the role may be related toregulation of LCA, LCA-induced liver injury in mice protection key genes SHP, of CYP7A1andCYP3A11’s. Conclusion: The cell-based screening experiments based on PXR in-of CYP3Apathway, we found that traditional Chinese medicine compound prescription and other applicationstaboo on the role of organisms in the regulation of CYP3A and the monomer composition is closelyrelated to: ginseng The reason and Veratryl contrary mainly due to the increase in the combinedtreatment, the dissolution of its monomer saponins, saponins are mostly inhibit cell expression ofCYP3A, so that the organisms on the metabolism of Veratryl major toxic components veratridinealkali, mediated Fen amine alkaloids. suppression. White peony in Siwutang corrigent effect "mayalso be with white peony can inhibit CYP3A expression correlation. Found a series of screeningexperiments on the monomer composition of the inhibition and induction potential of compoundsmonomer obtained by the conformation of the monomer components and structure of the structureanalysis of the molecular conformation of CYP3A4induction and PXR receptor binding capacityhave a decisive impact. At the same time, the use of excess lithocholic acid caused liver injurymodel provides a good basis for the work to solve the bile acid deposition in the liver injury causedby, in order to further investigate the potential ability of the compounds induced CYP3A4monomercomposition in the role of the liver detoxification.