Comparative Serum Metabolomic Study Of Pinaverium Bromide And Anchang Decoction Intervention In Rat Model Of IBS-D

Objective: In this study,we used two different therapeutic drugs,Anchang Decoction and pinaverium bromide,to intervene in rat model of irritable bowel syndrome(IBS)with liver depression andspleen deficiency syndrome.The serum metabolite spectral in rats were detected by using liquid chromatography-mass spectrometry(LC-MS),we screened the differential metabolites,deduced the relevant metabolic pathways involved in each,compared the similarities and differences in their effects on the metabolic patterns of rats with irritable bowel syndrome,and provided possible metabolomic explanations.Methods: 48 male Sprague-Dawley rats(SPF)were randomly and equally divided into normal group,model group,Pinaverium bromide group,and Anchang Decoction groups.The IBS-D rat model was constructed by capsaicin gavage combined with restraint stress.The visceral sensitivity and fecal condition of rats were assessed using abdominal withdrawal reflex and diarrhea index.After successful modeling,serum samples of rats in each group were collected,and serum metabolites of rats in each group were detected by LC-MS,combined with univariate statistical analysis and multivariate statistical analysis methods such as PCA and OPLS-DA to filter the differential metabolites,and to obtain the significant differential metabolites between compared groups.Pathway enrichment analysis of differential metabolites was based on the KEGG database,and the possible metabolic mechanisms of Anchang Decoction and pinaverium bromide for IBS-D were discussed based on the results.Results: The observation of the general state of the rats showed that compared with the normal group,the rats in the other groups after modeling by gavage of capsaicin combined with restraint stress showed dark hair color,hair loss,decreased activity,decreased water consumption and appetite,mental depression,increased number of loose stools and defecation frequency.The results of scoring the abdominal withdrawal reflex showed that the visceral sensitivity of the rats increased after modeling compared with the normal group(P<0.05).And the results of measuring the diarrhea index of the rats showed that the diarrhea index of the rats increased after modeling compared with the normal group(P<0.05).Modeling is successful.The results of metabolomics data analysis showed that there were50 differential metabolites in the Anchang Decoction group compared with the model group,of which 16 metabolites showed an upward trend in content and 34 metabolites showed a downward trend in content.There were 38 different metabolites in the Pinaverium bromide group compared with the model group,of which 12 metabolites were up-regulated and 26 metabolites were down-regulated.The results of metabolic pathway enrichment analysis showed that the possible metabolic pathways involved in Anchang Decoction are:valine,leucine and isoleucine biosynthesis,tyrosine metabolism,phenylalanine metabolism,tryptophan metabolism,fructose and mannose metabolism,primary bile acid biosynthesis,butanoate metabolism,synthesis and degradation of ketone bodies,retinol metabolism.The possible metabolic pathways involved in pinaverium bromide are: valine,leucine and isoleucine biosynthesis,tyrosinemetabolism,phenylalanine metabolism,Inositol phosphate metabolism,pyrimidine metabolism,ascorbate and aldarate metabolism.Conclusion: Both pinaverium bromide and Anchang Decoction can affect the metabolic profile of IBS-D rats,and among them valine,leucine and isoleucine biosynthesis,tyrosine metabolism,and phenylalanine metabolism may be a similar metabolic pathway for pinaverium bromide treatment and Anchang Decoction treatment for IBS-D.The release of intestinal spasm by pinaverium bromide may be associated with the Inositol phosphate metabolism.In addition,Anchang Decoction may also treat IBS-D by modulating the butanoate metabolism,Tryptophan metabolism and Primary bile acid biosynthesis.