| Objective:To prepare a rat model of diarrhea-predominant irritable bowel syndrome by acetic acid enema and bundle binding method,and to treat diarrhea-predominant irritable bowel syndrome rats with herb-partitioned moxibustion and investigate the effects of herb-partitioned moxibustion on intestinal flora and metabolites.Methods:Forty male SPFSD rats were divided into control group(12 rats)and model group(36 rats)after one week of adaptive feeding.The rats in the model group were subjected to acetic acid enema for 7 days and then restrained stress for 2 weeks.Rats in the original model group were randomly divided into a model group,a western medicine group and herb-partitioned moxibustion group,with 12 rats in each group.The rats in the normal groupwas givenno intervention.The rats in model group was bundled and not treated.The rats in herb-partitioned moxibustion group was treated with herb-partitioned moxibustion treatment,and The rats in western medicine group was given pimevir bromide tablet by gavage.After 2 weeks of treatment,the shape of Bristol stool was scored.The efficacy was evaluated in terms of fecal water content and visceral sensitivity evaluation.Two stool samples were collected using a sterile EP tube for metabolomics and intestinal flora testing.Based on Illumina high-throughput sequencing platform,the changes of intestinal flora abundance and diversity between rats in each group were studied,and the recovery of intestinal flora after intervention of herb-partitioned moxibustion was analyzed by LEfSe analysis.Metabolomics studies were performed on stool samples from rats in each group using nuclear magnetic resonance imaging.The nuclear magnetic resonance spectra were pre-processed and normalized by MestReNova software,and imported into SIMCA-P14.1 software for multi-dimensional statistical analysis.The characteristic metabolitesin rat body fluids were screened by VIP values in the OPLS-DA model combined with t-test.Through biological function analysis,find related metabolic pathways.Results:1.The fecal trait score and fecal water content of the model group were higher than those of the normal group,and the AWR score was 3 points.The water injection volume was significantly lower than that of the normal group(P<0.01),and it returned after the intervention of the herb-partitioned moxibustion.It is indicated that the herb-partitioned moxibustion has aregulation effect on diarrhea and visceral hypersensitivity in the IBS-D rat model prepared by acetic acid enema and restraint stress.2.The results of α diversity analysis showed that the effect of herb-partitioned moxibustion on the diversity and abundance of intestinal flora was obvious.ANOSIM similarity analysis and β diversity analysis showed significant differences in intestinal flora between the model group and the control group.The herb-partitioned moxibustion can adjust the content of Akkermansia muciniphila,Prevotella,Bacteroidetes to restore normal.3.Metabolite changes in the model group includedthe increased sucrose and uridine diphosphate glucose concentrations and decreased leucine,isoleucine,acetic acid enema,proline,2-hydroxybutyrate,valine,lactic acid,ethanol,arginine,2-oxoisovalerate and bile acid.They were mainly involved inthree major metabolic pathway including sarginine and proline metabolism,propionate metabolism and degradation of proline,leucine and isoleucine.After the intervention of herb-partitioned moxibustion,the metabolic state of the disordered rats in the IBS group can be restored to a normal state to some extent.Conclusion:1.The herb-partitioned moxibustion had a regulation effect on the diarrhea symptoms and visceral hypersensitivity state of the IBS-D rat model prepared by acetic acid enema and restraint stress.2.The herb-partitioned moxibustion has a good regulation effect on the diversity of IBS-D intestinal flora.Herb-partitioned moxibustion can regulate some of the flora,and affect the mucin degradation,the intestinal barrier function,and the inflammatory reaction.3.Herb-partitioned moxibustion can restore the metabolic disorder of IBS-D rats and affect immune response,intestinal barrier integrity,intestinal epithelial cell inflammation,and bile acid reabsorption to improve the symptoms of IBS. |
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