| BackgroundDiarrhea is one of the main causes of death among children under 5 years old.The world's statistics account for fifth.In China,the incidence rate of diarrhoea is about 5 per year.Among them,the proportion of children under5 years old is more than half.Diarrhea in children is a global public health problem.Qiweibaizhu decoction(QBD)is a classical prescription for the treatment of diarrhea with spleen deficiency syndrome.In recent years,a large number of pharmacological studies of traditional Chinese medicine(TCM),clinical and animal experimental studies have confirmed its efficacy and part of the mechanism of diarrhea,but its mechanism is not fully clear at present.At present,the relationship between intestinal flora,short chain fatty acids(SCFAs),intestinal mucus layer and diarrhea has been confirmed by continuous research.Diarrhea and intestinal flora disorder are mutual cause and effect.Any kind of diarrhea is accompanied by intestinal flora disorder.Intestinal flora disorder can also lead to and aggravate diarrhea.SCFAs come from the digestion of carbohydrate in the intestinal tract by intestinal flora,which can stimulate the growth of colonic epithelial cells Na + depends on the absorption of water and electrolyte to alleviate diarrhea symptoms.Mucin2(MUC2),the main protein in the intestinal mucus layer,is an important component of the intestinal mucus barrier.It plays an important role in the defense of pathogenic microorganisms and maintaining the balance of intestinal flora.It is worth noting that in recent years,studies have shown that the increase of MUC2 content helps to alleviate diarrhea.ObjectiveCombined with alcian blue staining and immunofluorescence laser confocal analysis,16 S r DNA amplicon sequencing analysis,GC-MS / MS detection and other technologies,taking QBD as the representative prescription,diarrhea rats with spleen deficiency syndrome as the experimental object,this paper studied on the metabolic mechanismthe of the intestinal microflora-SCFAs in the treatment of diarrhea with spleen deficiency syndrome in children by syndrome differentiation,and the mechanism of the intestinal microflora-intestinal mucus layer and MUC2.MethodsForty-eight specific-pathogen-free male Sprague-Dawley rats(age,5weeks;weight,142±8 g)were randomly divided into three groups: the control group,the model group,Saccharomyces boulardii(S.boulardii)group and QBD group.The control group did not receive any treatment,the other three groups recieved polyethylene glycol(PEG 3350)to create the diarrhea model of juvenile rats,and the spleen deficiency syndrome model was made by using the small platform where juvenile rats would fall into the water when sleeping and had to consume physical strength to stand or swim continuously;then,the S.boulardii group and the QBD group were treated by oral administration of S.boulardii and QBD respectively,and the model group was treated by oral administration of 0.9% Na Cl solution.The feces,body weight and mental activity of the juvenile rats in each group were observed and recorded regularly every day.The juvenile rats were killed within 24 hours after treatment with corresponding drugs according to the course of treatment.The colon and colon contents of the juvenile rats were taken for detection.The expression and distribution of MUC2 and MUC2 in the colon were evaluated by alcian blue staining and immunofluorescence laser confocal analysis;the bacterial diversity and structural changes in the colon contents of juvenile rats were detected by 16 S r DNA amplicon sequencing;the content of SCFAs in the colon contents of juvenile rats was detected by GC-MS / MS.Results1.QBD treatment has the increasing tendencies of the weight of diarrhea rats with spleen deficiency syndromeCompared with the other three groups,the body weight of the control group had significant statistical differences;compared with the model group,there was no significant difference in the body weight of the S.boulardii group and the QBD group,However,the mean weights of the three groups were in the following order: QBD group> S.boulardii group> model group.2.QBD treatment tends to repair the mucus layer and promote MUC2 expression in diarrhea rats with spleen deficiency syndromeThe results of alixin blue staining showed that the diarrhea caused by PEG 3350 lead to the colon tissue of juvenile rats incomplete,and the mucus layer was significantly thinned or even missing.Only a few mucus were secreted from the basal part of the goblet cells,and most goblet cells did not secrete new mucus.QBD could improve the damage caused by PEG 3350.Immunofluorescence laser confocal analysis showed that the expression of MUC2 was low in the model group,while QBD could increase the expression of MUC2,which was consistent with the result of alixin blue staining.3.QBD treatment tends to increase the propionic acid concentration in diarrhea juvenile rats with spleen deficiency syndrome,but the effect is not as good as that of s.boulardii powder,and s.boulardii can also increase the concentration of acetic acid and butyric acid,suggesting that s.boulardii may have a more obvious effect on the concentration of SCFAs in diarrhea juvenile rats.4.QBD treatment can regulate the intestinal flora of diarrhea rats with spleen deficiency syndromePEG 3350 significantly changed the diversity and structure of intestinal flora in juvenile rats.Although QBD did not significantly change the diversity and structure of intestinal flora in juvenile rats,it enhanced the increasing tendencies of Verrucomicrobia and Akkermansia.Moreover,the S.boulardii significantly increased the abundance of Parasutterella.ConclusionsQBD has the tendency to improve the weight growth of diarrhea rats.QBD has a stronger ability to repair the mucus barrier than S.boulardii,but its ability to improve SCFAs is not as good as that of S.boulardii.S.boulardii may play a role in the treatment of diarrhea by regulating the intestinal flora to increase SCFA production,and the mechanism of action of QBD in the treatment of juvenile rats with diarrhea likely involves gut microbiota-mediated repair of the mucus barrier. |
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