The Absorbed Constituents And Metabolism Study Of The Chemical Components From Desmodii Styracifolii Herba In Rats

Objective:A high performance liquid chromatography tandem mass spectrometry method was used to analyse the discrepancies of in vivo and in vitro metabolic profiling of isovitexin under the normal and nephrolithiasis pathological conditions,and the absorbed constituents and metabolism study of the chemical components from Desmodii Styracifolii Herba?DSH?in kidney stone model rats was conducted.Methods:1. Animal model of calcium oxalate kidney stone was established orally by administration of 2%?w/v?ammonium chloride?1.0 m L/100 g?and 1.25%?v/v?ethylene glycol?EG?in drinking water.After oral administration of isovitexin to normal and kidney stone model rats,the bio-samples were collected.The UHPLC-Q-TOF-MS/MS technology was developed to acquire the data of bio-samples from in vivo and in vitro.Metabolitepilot^TM 2.0software was used to analyse and identify the existent metabolites.The discrepancies of metabolites between normal and kidney stone model rats were compared?2.Then,the extract of DSH was achieved by ethanol and orally administrated to kidney stone model rats.The data of biosamples were obtained by the method of UHPLC-Q-TOF-MS/MS.The absorbed components and metabolites of flavonoids from the DSH extract in the kidney stone model rats were analyzed by Metabolitepilot^TM 2.0 software.In addition,the molecular network analysis strategy was used to further identify the absorption components of DSH in kidney stone model rats.Results:After oral administration of isovitexin,18 phase I and 10 phase II metabolites were identified in normal rats,while 20 phase I and 13 phase II metabolites were detected in the kidney stone model rats.The results of relative quantitative determination reflected that the amounts of deglycosylation,reduction metabolites in the kidney stone model rats were much more than those in normal rats.Instead,the levels of the oxidation and dehydrogenation metabolites were conversely higher in the normal samples compared with the kidney stone model samples.When the extract of DSH was lavaged to the kidney stone model rats,221 metabolites of flavonoids were identified in biosamples,which 16 prototype components and 82 metabolites of flavonoids were detected in serum.The main metabolic pathway was hydrogenation,oxidation,hydrolysis,deglycosylation,methylation and glucuronide conjugation.A total of 49 prototype components including 45flavonoids and 4 phenolic acids were identified based on molecular network analysis of absorption components of DSH in serum samples.Conclusion:1.The eatablished UHPLC-Q-TOF-MS/MS method can accurately and rapidly identify the metabolites of isovitexin,which provides a theoretical basis for further study on the metabolism of the total extract of DSH and shows that the study of drug metabolism with pathological model animal is more practical in clinical practice.2. The metabolic situation of the DSH extracts in kidney stone model rats was studied in the first time,and the absorbed components and metabolites of flavonoids in the DSH extracts were analyzed.This experiment laid a foundation for determining the pharmacodynamic substances of DSH,and further provides a basis for the the determination of its quality markers.