| In traditional Chinese medicine(TCM),an "herb couple" is made up of two relatively fixed herbs paired according to their compatibility and the theory of "Qi Qing He He"(the harmony of seven affects,or emotions).The Chinese herbal property theory describes the herb nature and character.Out of many properties,heat and cold is the most important herb property.Huang Lian(Rhizoma Coptidis,HL)and Wu Zhu Yu(Fructus Evodiae,WZY)are respectively a cold and a hot herb,and they are paired based on the "heat/cold couple"principle.Since coptis and Fructus Evodiae have opposite properties,different coupling ratios may lead to varying drug efficacy.However,present studies of coptis and Fructus Evodiae found that,without being coupled,coptis and Fructus Evodiae still exert a therapeutic effect on both Stomach Heat SyndromeC SHS)and Stomach Cold SyndromeC SCS),and biochemical indicators failed to reflect the change of herb properties when coptis and Fructus Evodiae are coupled at different ratios.Metabolomics,on the other hand,is a holistic research method based on a more comprehensive view,which is consistent with the "overall" and "dialectical"principles of TCM.Thus,this study used a metabolomic method to investigate the influence of coptis/Fructus Evodiae ratios on drug properties,and to discuss the scientific connotation of the concept“to heat the cold,and to cold the heat".This study used SHS and SCS rats as pathological models,and used a metabolomic method to compare and evaluate the efficacy of coptis and Fructus Evodiae in different pathological models.The main contents are as follows:Metabolomic study of rat models with SHS and SCSUsing GC-MS-based plasma metabolomics to study metabolism changes in rats with SHS and SCSBy using principal component analysis,this study found that the metabolites of normal,SHS and SCS rats were clustered into three obvious groups,indicating that under different physiological states,rats will show significantly different expression of metabolites.Further,we used PLS-DA to analyze and screen for biomarkers associated with SHS or SCS.According to the resuLt,36 plasma biomarkers were identified to be associated with SHS,and 16 were associated with SCS.Noticeably,though some biomarkers for SHS overlapped with those for SCS,most biomarkers were non-identical.Also,even if different syndromes influenced the same biomarker,they show different reguLating directions.On the whole,these biomarkers couLd be used to distinguish SHS from SCS.Then,this study used intelligent metabolic pathway analysis to identify metabolic pathways that are associated with SHS and SCS.The resuLts showed that,SHS mainly affected the synthesis of alanine,tyrosine and tryptophan,and the metabolism of phosphoinositide,tyrosine,arginine,proline,and anabolic steroids.On the other hand,SCS mainly affected the metabolism of alanine,aspartic acid,glutamic acid and glycerol phospholipid.Using UPLC-QTOF-MS-based plasma metabolomics to study the overall metabolic changes in SHS ratsPrincipal component analysis found that the metabolites of normal and SHS rats were clustered into two clear groups,indicating that rats under different physiological conditions showed significantly different expression of metabolites.Further screening identified 10 plasma biomarkers associated with SHS,and intelligent metabolic pathway analysis found that glycerol phospholipid metabolism might be most closely related to SHS.Data fusion-based metabolic pathway analysis of rat SHS modelSHS metabolomics data obtained from UPLC-QTOF-MS and GC-MS were fused by the muLti-block PCA algorithm,and principal component analyze showed that normal and SHS rats were clustered into two clear groups,indicating that rats under different physiological conditions showed significantly different expression of metabolites.From the load diagrams of all blocks,47 plasma biomarkers were identified to be associated with SHS,and most of them were overlapped with those identified by GC-MS and UPLC-QTOF-MS analysis.Then,metabolic pathway analysis was carried out to further identify SHS-associated metabolic pathways,and the resuLt showed that SHS mainly influenced the synthesis of phenylalanine,tyrosine and tryptophan,and the metabolism of phenylalanine,glycerol phospholipid,glyoxylic acid,phosphoinositide,tyrosine,arginine,proline and anabolic steroids,as well as the citric acid cycle(TCA cycle)metabolic pathway.Mechanisms by which coptis and Fructus Evodiae affect drug propertiesGC-MS-based metabolomic analysis of the influence of coptis/Fructus Evodiae herb couples on SHSBased on the TCM theory“to cold the heat,and to heat the cold",the efficacy of coptis-Fructus Evodiae couple 6:1(Zuo Jin Wan,ZJW),theoretically,exceed that of coptis-Fructus Evodiae couple 1:6(Fan-Zuo Jin Wan,FZJ).In this study,we found that both ZJW and FZJ improved SHS.However,according to the PCA score plot,clustering of metabolites in rats treated with ZJW was much closer to that of the normal group,while the cluster in rats treated with FZJ was more similar to the model group.ZJW interfered with five metabolic pathways significantly associated with SHS,and the levels of related metabolites,i.e.L-tyrosine,inositol-1-phosphate,pyruvate,L-phenylalanine,campesterol and cholesterol were restored to normal.On the other hand,FZJ affected three metabolic pathways,and significantly reguLated the levels of related L-tyrosine,cholesterol and campesterol.These resuLts indicate that the coupling of coptis and Fructus Evodiae is in line with the theory of"to cold the heat,and to heat the cold".Our study found that both coptis and Fructus Evodiae improved SHS to different levels,but the PCA score plots showed that the duster of coptis was closer to the normal group while the Fructus Evodiae was closer to the model group.Considering the comparison of ZJW and FZJ,this study concludes that both coptis and Fructus Evodiae have a therapeutic effect on SHS,but coptis is the main active ingredient.Coptis and Fructus Evodiae display both synergistic and antagonistic interactions in treating SHS.Regarding to the SHS model group,we also compared the change of metabolic profilebefore and after berberine intervention.According to the PCA plot,the Fructus Evodiae group was clustered farthest from the normal group,and the berberine group was clustered in partial overlap with the coptis group.This suggests that berberine and coptis may have similar effects on SHS,and further indicates that in one aspect,berberine is the active ingredient of coptis in treating SHS,and in another aspect,berberine also has a "heat" drug property.UPLC-QTOF-MS-based metabolomic analysis of the effect of coptis/Fructus Evodiae herb couples on SHSOut of the biomarkers for SHS,10 were selected and analyzed,i.e.,propionic agmatine,adrenal acid,LysoPC(18:1),LysoPC(18:2),LysoPE(20:0),PS(18:1),PS(19:0),PC(19:0),PS(20:1)and LysoPC(16:0).These metabolites are associated with the synthesis and release of arachidonic acid,which is an important precursor for eicosanoids during the inflammatory process.The treatment of SHS by ZJW,FZJ,coptis and Fructus Evodiae may possibly depend on restoring the disorders of the glycerol phospholipid metabolism pathway.This study compared and analyzed the metabolic profile before and after coptis and berberine intervention.The PCA plot suggests that berberine and coptis may have similar mechanisms in treating SHS,indicating that in one aspect,berberine is the active ingredient of coptis intreating SHS,and in another aspect,berberine also has a "heat" drug property.GC-MS-based metabolomic analysis of the effect of coptis/Fructus Evodiae herb couples on SCSTheoretically,the treatment efficacy of FZJ on SCS shouLd be better than ZJW.However,in this study,we found that when compared with FZJ,clustering of ZJW tended to be deviated For example,the reguLation of benzoic acid by ZJW and FZJ both tended to cluster towards the model group,but ZJW showed more significant deviation.Comparing with the normal,the model group had lower levels of 4-hydroxybutyric acid,2-amino butyric acid,capric acid and myristic acid.In ZJW and FZJ groups however,the level of these metabolites were increased,but the ZJW group showed levels significantly higher than those of the normal,indicating its cluster was deviated.Besides,as for the reguLation of 1H-indole-2,3-dione,glutamine,glycerol-3-phosphate and ribonic acid,the cluster of FZJ was closer to the normal while that of ZJW was closer to the model.This also indicates that the properties of coptis and Fructus Evodiae are in line with the theory of“to cold the heat,and to heat the cold".When compared between coptis and Fructus Evodiae,we also found that clustering of coptis was prone to deviation.For example,the reguLation of D-galactose-1,4-lactone by coptis and Fructus Evodiae both tended to cluster towards the model,but coptis showed more significant deviation.When compared with the normal,the model group showed lower levels of capric acid and myristic acid.Both coptis and Fructus Evodiae elevated the expression of these metabolites,but coptis significantly increased these metabolites to a level even higher than the normal,indicating that the cluster of coptis was deviated.Besides,regarding to the reguLation of 4-hydroxybutyric acid and palmitoleic acid,Fructus Evodiae clustered closer to the normal while coptis clustered closer to the model.This again suggests that the properties of coptis and Fructus Evodiae are in line with the theory of "to cold the heat,and to heat the cold".Chemical composition of Herb property Huang Lian Wu Zhu Yu and endogenous metabolites association studyOf which 22 endogenous metabolites study,there are no significant difference between group HL and ZJW,suggesting the content of HL and ZLW changed little in the chemical composition,endogenous metabolites influence is insignificant;there were no significant differences between groups FZJ and WZY,suggesting the content of FZJ and WZY changed little in the chemical composition;there are no significant difference between group ZJW and normal;there are no significant difference between groups FZJ and model;there are significant difference between groups FZJ and ZJW.Chemical composition and endogenous metabolites association studies can completely characterize the substance ZJW and FZJ potency differences.This indicates that for the treatment of SHS,ZJW is in line with the theory of“to cold the heat".This shows that the treatment of stomach disorders,in order to comply with TCM herbsZuojinwan "heat of those cold" theory..Then,this study used intelligent metabolic pathway analysis to identify metabolic pathways that are associated with ZJW and FZJ.The resuLts showe phenylalanine,tyrosine and tryptophan biosynthesis,D-glutamine,and D-Metabolism of glutamate metabolism,p-alanine metabolism,metabolism of phenylalanine,arginine and proline metabolism,alanine,aspartic acid and glutamic acid metabolism,phosphoinositide metabolism,pyruvate,tyrosine metabolism,glycolysis or gluconeogenesis,glutathione metabolism,citric acid cycle(TCA cycle)pathways are effected.Ethanol-induced gastric ulcer model as an exploratory study using stomach Syndrome and its applicationUsing rats with ethanol?induced acute gastric mucosal damage as SHS model,the therapeutic effects of coptis,Fructus Evodiae and different coptis-Fructus Evodiae couples were evaluatedOur study found that,coptis-Fructus Evodiae couple 6:1(Zuo Jin Wan,ZJW)and coptis-Fructus Evodiae couple 1:6(Fan-Zuo Jin Wan,FZJ)exerted different levels of protective effects on ethanol-induced rat acute gastric mucosal damage.Out of the tested drugs,ZJW had the best efficacy while Fructus Evodiae showed no significant effect,indicating that coptis is the main active ingredient of the herbal couple when treating SHS.Indeed,this resuLt is consistent with the herbal property theory that coptis-based couples have a cold property,mainly function to purge fire,and are able to cure SHS.Our study also found that,the main mechanisms by which coptis/evdia couples treat SHS are anti-oxidation and anti-inflammation.Our study found that,coptis-Fructus Evodiae couple 6:1(Zuo Jin Wan,ZJW),coptis-Fructus Evodiae couple 1:6(Fan-Zuo Jin Wan,FZJ)and coptis exerted different levels of protective effects on ethanol-induced rat acute gastric mucosal damage.Out of the tested drugs,ZJW had the best efficacy while Fructus Evodiae showed no significant effect,indicating that coptis is the main active ingredient of the herbal couple when treating SHS.Indeed,this resuLt is consistent with the herbal property theory that coptis-based couples have a cold property,mainly function to purge fire,and are able to cure SHS.Our study also found that,the main mechanisms by which coptis/evdia couples treat SHS are anti-oxidation and anti-inflammation.Comparison of therapeutic effects of main alkaloids in coptis on SHSThis study further compared the protective effects of main alkaloids in coptis on ethanol-induced rat acute gastric mucosal injury.The resuLts suggested that berberine suLfate exerted a stronger mucosal-protective role when compared with berberine hydrochloride.In coptis,berberine suLfate,palmatine,jatrorrhizine and coptisine all show varying degrees of protection against gastric mucosal injury,but berberine suLfate and palmatine showed the highest efficacy,and these two substances protected the mucosa mainly by anti-oxidation and anti-inflammation.Comparing the therapeutic effects of berberine/saponins couples on SHSThis part is designed for the development of new natural drugs or health products that can prevent or relieve gastric uLcer,and are with low toxicity and few side effects.In this study,the protective effects of berberine,saponins(from Momordica charantia L.leaves),and berberine/saponins couple were compared in rat with ethanol-induced acute gastric mucosal damage.The resuLt showed that,the berberine/saponins couple showed a stronger protective effect when compared with berberine and saponins. |
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