Studies On Absorption,Transport And Metabolism Of Lignans In S.chinendsis Based On In Vitro Model

The dry and rape fruits of Schisandra chinensis?S.chinensis?are used as a kind of nourishing traditional Chinese medicines with the functions of inducing astringency,replenishing and promoting production of body fluid and tonifying the kidney to relieve mental strain.This paper studied the absorption and transportation mechanism of lignans in S.chinensis in vitro by applying Caco-2 cells and MDCK cell models for simulating the absorption process of the small intestine,combined with the technology of high-performance liquid chromatograpHy?HPLC?and mass spectrometry;In vitro enterobacteria and liver microsomal metabolism models were used to investigate the rule of metabolism of lignans in S.chinensis.Firstly,MDCK monolayer cell model was applied to investigate the absorption and transportation characteristics of lignans from extraction and purification of S.chinensis,which includes Schisandrin A,Schisandrin B,Schisandrin C,S chisandrol A,Schisandrol B,Schisantherin A,Schisantherin B and their corresponding standard products.The experiment investigated the effect of time on the transportation of these seven kinds of schisandrin lignans in the MDCK monolayer cell model.The sample concentration was detected by the liquid-mass spectrometry,and the apparent permeability coefficient?Papp?and overflow ratio?Er?were used for experiment evaluation.The results showed that the Rapp of the six major lignans in the S.chinensis,except for Schisandrin C,were greater than 1×10^-6 cm/s,indicating the characteristics of the good oral absorption.The degree of cellular absorption is higher in Schisandrol A and Schisandrol B,followed by Schisantherin A and Schisantherin B,while Schisandrin A and Schisandrin B showed a lower degree than the former lignans.In addition,the Er value of these six lignans are between 0.7 and 1.2,indicating that the absorption process in the gastrointestinal tract is mainly passive transportation.Secondly,using the two in vitro absorption models of MDCK cells and C aco-2 cells,the absorption and transport mechanisms of fifteen lignans in Schisandra chinensis extract and lignan purified were studied.The results show that all lignans except Gomixin E are well absorbed.It is preliminarily concluded that Gmisin D is a P-glycoprotein substrate,and its Rapp and Er value are greater than 1.5,which is furtherly confirmed by the P-glycoprotein inhibitor verapamil.The lignans are better absorbed in Caco-2 cells than MDCK cells,indicating that the absorption carrier protein in Caco-2 cells promotes the absorption of lignans in Schisandra chinensis.Thirdly,using in vitro metabolic model of enterobacteria,we carried out in vitro metabolism study of Schisandra lignan.The experiment established an in vitro enterobacterial metabolism model of healthy mice,healthy rats and AD rats,combined with liquid-mass spectrometry technology,to extract Schisandrin A,seven Schisandra mixed standard products,Schisandra lignan purified products and Schisandra extract Detection and analysis of metabolites.The results showed that S.chinensis remains stable among the gut microbiota in rat,and the gut microbiota does not metabolize lignans.Fourthly,the in vitro liver microsome metabolism model was applied to investigate the in vitro metabolism of the extracted and purified products of S.chinensis.The main oxidase and enzyme in the hepatic microsome cytochrome P450 enzyme system is the liver microsome.Oral drugs are absorbed into the intestinal villi epithelial cells to enter the body,and then undergo an enzyme metabolism reaction through the liver before exerting further pHarmacological effects in the systemic circulation to.In the experiment,the in vitro liver microsome incubation model was used to study the liver microsome metabolic behavior of S.chinensis,and UHPLC-Q-TOF-MS technology was used for concentration detection.The results showed that S.chinensis can remain stable in the sliver microsomal P450 enzyme system without being metabolised.This paper studied the absorption and transportation mechanism of lignansin S.chinensis in vitro by applying Caco-2 cells and MDCK cell models for simulating the absorption process of the small intestine,combined with the technology of high-performance liquid chromatograpHy?HPLC?and mass spectrometry;In vitro enterobacteria and liver microsomal metabolism models were used to investigate the rule of metabolism of lignans in S.chinensis.This study aims at laying a foundation for the study of the mechanism of pHarmacological effects of lignans in S.chinensis,and also provides scientific evidence for their clinical application.