| With the advent of the great health era,the inheritance and development of Chinese medicine have ushered in a new dawn.Faced with unprecedented opportunities and challenges,it's no doubt that the development of Chinese medicine plays a pivotal role in it.Chinese medicine decoction piece is playing a more prominent role in clinic of Chinese medicine.But the theory of Chinese herbal medicine processing which guide its rational application is still a lack of modern scientific arguments.Clinical drug mixed,chaotic,miscellaneous phenomenon still exist and have become a bottleneck restricting the development of Chinese medicine.Traditional Chinese medicine processing is a profound subject.Not only ingredients,but also efficacy,drug properties and channel tropism have been changed while processing.It is inappropriate to determine the quality of herbs according to several chemical components.This subject with Schisandrae chinensis Fructus explores the internal mechanism systematically and comprehensively from the following three aspects:Firstly,The change of material basis through the processing.Secondly,The difference of total lignans in Schisandrae chinensis Fructus which happens during metabolism in rats.Thirdly,The metabolomics research of Schisandrae chinensis Fructus against alcohol-induced liver injury.Thus,we can provide scientific theoretical basis for the study of traditional Chinese medicine processing.The experiment is divided into three parts:1)Screening of potential chemical markers before and after processing of Schisandrae chinensis Fructus.2)Internal qualitative analysis of alcohol extracts of Schisandrae chinensis Fructus.3)Metabolomics study on the effects of crude and vinegar-processed Schisandrae chinensis Fructus on anti-alcoholic liver injury.1.Screening of potential chemical markers before and after processing of Schisandrae,chinensis Fructus In this research,we established a method of combining UPLC-Q-TOF/MS with multivariate statistical analysis to filtrate the potential chemical composition rapidly,that is chemical markers.Then validate these potential chemical markers by mass spectrometry,retention time,and/or tentative assignment of empirical formula by reference to reference compounds in the literature or databases.by mass spectrometry,retention time,and/or tentative assignment of empirical formula by reference to reference compounds in the literature or databases.These potential chemical markers are not only used to distinguish between Chinese medicinal herbs and processed products,but also may be a component of the Chinese herbal medicine that is the basis of effective substances.Using the method mentioned above,this study screened 40 different components of Schisandrae chinensis Fructus after vinegar processed,among which,there were 8 significant differences chemical composition among the crude and vinegar-processed Schisandrae chinensis Fructus(chemical markers).Five chemical constituents were identified,they are 5-hydroxymethylfurfural(5-HMF),schisandrin A and its isomer,schisandrin B and schisantherin D,respectively.The other 3 chemical markers,by analyzing the MS and MS/MS spectra,speculate that they are likely to belong to lignans.Among them,the schisandrin B and 5-HMF have very significant differences,and have been identified as the most representative chemical markers.To sum up,the chemical composition of Schisandrae chinensis Fructus after vinegar processed has undergone a series of complex changes,especially lignans.Therefore,it is speculated that lignans are the important material basis for hepatoprotective effect of vinegar processed Schisandrae chinensis Fructus.2.Study the on metabolism of liver injury in rats of crude and vinegar-processed Schisandrae chinensis Fructus According to the research,the main substances of extractive of Schisandrae chinensis Fructus are lignans.By the verification of reference substance and the quantitative analysis,Deoxyschizandrin,Schisandrin B,Schisandrin C,Schizandrol,Schizandrol B,Schisantherin,Schisantherin B,Schisanhenol,Gomisin J,Gomisin G etc.are found in lignans.The front eight lignans element content is higher.In recent years,studies on the metabolites of lignans in Schisandrae Chinensis Fructus are also limited to the monomer composition.It is hard to see specific report that is concerning extract the metabolism of the body after dosing study.To study traditional Chinese medicine(TCM)in the body's metabolic product,can reverse identification the substances that are absorbed or have curative effect,greatly accelerate the determination of effective components group and further reveal the material basis of drug action and mechanism of action.This study uses UPLC-Q-TOF/MS technology,and the quality loss of filtration(MDF)method,a rapid search for target metabolites in complex matrices of biological samples of Chinese medicine was successfully achieved,and comparative studies on the metabolites of lignans from the extracts of Schisandra chinensis and vinegar were carried out in rats.The results showed that lignans mainly occurred such as removal of methoxyl group,methylene and oxidation reaction in vivo.Compared with the prototype compounds,the retention time of the corresponding metabolites was shortened,that is,polarity increased.The vast majority of lignan metabolites are distributed in crude and vinegar Schisandrae Chinensis Fructus group's plasma,bile,urine or feces samples,however the metabolites in vinegar Schisandrae Chinensis Fructus group except M18,the remaining in the bile have higher distribution and content,but the metabolites in crude Schisandrae Chinensis Fructus group nearly half of them were not detected or response too lower in bile.After vinegar processing,Schisandrae Chinensis Fructus really have changed the composition ratio of total lignans,which led to the absorption,distribution and metabolic differences of lignans.The metabolites of lignans in the vinegar-processed Schisandrae Chinensis Fructus group were significantly higher in the bile and feces than in the crude Schisandrae Chinensis Fructus group.The distribution of lignan metabolites in the bile and feces,suggesting that its mechanism may be related to the enterohepatic circulation and intestinal microflora.Through the above research,it is expected to provide an important basis for further exploring the material basis and mechanism of vinegar-processed Schisandrae Chinensis Fructus enhancing the hepatoprotective effect.3.Metabolomics study on the effects of crude and vinegar-processed Schisandrae Chinensis Fructus on anti-alcoholic liver injury The liver is the center of body's substance metabolism,Its metabolic transformation and phagocytosis function has an important protective effect on the body.At present,liver disease caused by various factors endanger human health seriously and it is one of the common problems facing the world today.The incidence rate of liver disease in our country is relatively high and its influence is a bit wide,the liver disease seriously endanger people's health and national economy.At present,most of the studies on the mechanism of Schisandrae Chinensis Fructus in the treatment of acute alcoholic liver injury have been carried out in terms of pharmacological effects and hepatic enzymes.From the perspective of metabolomics,there are few reports on the effects of vinegar on Schisandrae Chinensis Fructus.The research group found that in the previous study,vinegar-processed Schisandrae Chinensis Fructus can promote bile secretion in rats,the total amount of bile excretion was significantly different from crude Schisandrae Chinensis Fructus.In this study,plasma,bile and urine are taken as the object of study to discuss the effects of drugs on liver system from the perspective of metabolomics.By using UPLC-Q-TOF/MS and pattern recognition,we aim to investigate the effects of crude and vinegar-processed Schisandrae Chinensis Fructus in metabolites in rat plasma,bile,urine group and excavate potential biomarkers to find possible mechanisms for the treatment of acute alcoholic liver injury with Schisandra.The purpose is to expound the scientific connotation of the effect of vinegar-processed Schisandrae Chinensis Fructus.Adopt the metabolic skills based on UPLC-Q-TOF/MS,PC A and OPLS-DA for the screening and identification of biomarkers related to liver injury.The Bile metabolite spectrum of normal control group,model group,crude Schisandra and vinegar-processed Schisandra group bile metabolite profile was significantly separated.We discovered and identified 21 differential metabolites in the plasma like PC,PG,PE,TG,Phytosphingosine,Palmitoyl Serinol etc.20 potential biomarkers related to liver injury LysoPC(20:4),PG(18:0/18:1),12-ketodeoxycholic acid,TG(64:2)etc in bile.Above all,it indicates that acute alcoholic liver injury in rats damages the lipid metabolism,bile acid metabolism and amino acid metabolism and other abnormalities.After giving crude and vinegar-processed Schisandrae Chinensis Fructus,the level of the above markers can be adjusted to the normal state,and the regulating function of the vinegar-processed one is stronger than that of the crude one(P<0.05).Vinegar-processed Schisandrae chinensis Fructus might have the anti-liver injury effect on all these metabolic pathways.4.This paper based on previous work,using UPLC-Q-TOF/MS technology combined with the method of multivariate statistical analysis.The changes of material basis before and after the processing of Schisandrae chinensis Fructus were analyzed,The metabolism and excretion of crude and vinegar-processed Schisandrae chinensis Fructus in rats were systematically analyzed,and tried to reveal the internal mechanism of vinegar-processed Schisandrae chinensis Fructus enhanced anti liver injury by metabonomics.The mechanism and function of vinegar-processed Schisandrae chinensis Fructus in protecting liver injury were discussed in terms of chemical composition,metabolic spectrum and metabonomics. |
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