| ObjectiveTo observe changes in the intestinal flora and intestinal mucosal barriers of rats after drug intervention,the traditional Chinese medicine professor Qing-sheng Li from Yunnan Province was used to treat the antibiotic-associated diarrhea effective prescription-Hetiao Jianpi Decoction.Initially clarified the mechanism of interfering antibiotic-associated diarrhea with Hetiao Jianpi Decoction,it provides theoretical basis for Chinese medicine to seek multi-target treatment of antibiotic-associated diarrhea,and provides new ideas for clinical prevention and treatment of the disease.Methods1.The rat model of antibiotic-associated diarrhea with spleen deficiency was established by giving two antibiotics and rhubarb decoction.The model was evaluated by rat behavioral changes and pathological changes of colonic tissue.2.Rats in each dose group of Hetiao Jianpi Decoction were administered with different concentrations of Hetiao Jianpi Decoction.The rats in the positi-ve control group were administered with Clostridium butyricum,and the rats in the control group and model group received physiological Intragastric saline.3.Rats plasma contents of diamine oxidase(DAO)、D-lactic acid(D-LA)and secretory immunoglobulin A(SIgA)in rat colon tissue by enzyme-linked immunosorbent assay.The expression of Toll-like receptor 4 mRNA in rat colon tissue was detected by Real-time fluorescence quantitative polymerase chain reaction.Rat intestinal contents short-chain fatty acids and intestinal contents abundance were respectively detected by gas chromatography and high-throughput sequencing of 16S rRNA genes.Results1.Rat colon histopathological section results:Compared with the model group,the myometrial and mucosal edema of the colon tissue of rats in the low,middle and high dose groups of Hetiao Jianpi Decoction and the positive control group were significantly improved,and the continuity of the intestinal mucosa was restored.It showed that the colonic tissue damage of AAD rats was significantly improved after treatment with Hetiao Jianpi Decoction.2.Rat intestinal mucosal barrier indicators results:(1)DAO and D-LA:①DAO:Compared with the model group,the plasma DAO expression in the low-dose group of Hetiao Jianpi Decoction was significantly reduced(P<0.01),and the middle-dose group showed a decreasing trend(P>0.05).The DAO expression in the high-dose group was significantly reduced(P<0.05).The expression of DAO in the positive control group showed a downward trend(P>0.05).②D-LA:Compared with the model group,the expression of D-LA was significantly decreased in the low,middle and high dose groups of Hetiao Jianpi Decoction and the positive control group(P<0.05).Combined with the changes in the content of DAO and D-LA in the plasma of AAD rats,it showed that the permeability of the intestinal mucosa of rats was reduced and the structural damage of the intestinal mucosa was repaired after treatment with Hetiao Jianpi Decoction.(2)SIgA and Toll4:①SIgA:Compared with the model group,The expression of SIgA in the colon tissue of the low-and middle-dose groups of Hetiao jianpi Decoction was significantly increased(P<0.05),and in the high-dose group showed an upward trend(P>0.05).In the positive control group,the expression of SIgA was significantly increased(P<0.01).②Toll4:The relative expression of Toll4 mRNA in the colon tissue of the low and high dose groups of Hetiao Jianpi Decoction was significantly reduced,which compared with the model group(P<0.01).The relative expression in the middle dose group was a downward trend(P>0.05).In the positive control group showed a downward trend(P>0.05).Combined with the SIgA content in the colon tissue of AAD rats and the transcription level of Toll4 mRNA,it was shown that the immune function of intestinal mucosa of rats was enhanced after treatment with Hetiao Jianpi Decoction.3.Rat intestinal flora results:(1)Relative abundance of intestinal flora:①Phylum level:The relative abundance of the four intestinal flora in the model group such as Bacteroidetes of the rats increased,while the relative abundance of the seven intestines of Firmicutes decreased.In the Hetiao Jianpi Decoction group,the relative abundance of the three intestinal flora such as Proteobacteria increased,while the two intestinal flora such as Spirochaetes decreased.In the positive control group,the relative abundance of the four phylums such as Firmicutes and Proteobacteria increased,and four such as Bacteroidetes decreased.②Genus level:In the model group,the relative abundance of ten bacterial genera such as Lactobacillus and Prevotella of the rat intestinal content flora increased,and the relative abundance of ten bacterial genera,such as Unspecified-Clostridiales and Unspecified-Lachnospiraceae decreased.The relative abundance of the four bacteria dominated by Unspecified-Rumin.ococcaceae and CF231 increased in the Hetiao Jianpi Decoction group,and the relative abundance of five bacteria dominated by Lactobacillus and Ruminococcus reduced-In the positive control group,the relative abundance of fourteen genus such as Lactobacillus and Unspecified-Clostridiales increased,while the six intestinal flora such as Unspecified-S24-7 and Prevotella decreased.Combined with the relative abundance changes of the intestinal flora phylum and genus levels of AAD rats,after treatment with Hetiao Jianpi Decoction,it showed that the structure of the intestinal flora of rats has changed.The number of beneficial bacteria increased,and pathogenic bacteria decreased.(2)SCFAs:Compared with the model group,the acetic acid and n-butyric acid contents in the high-dose Hetiao Jianpi Decoction group showed an upward trend(P>0.05).The content of propionic acid in the low-and high-dose groups of Hetiao Jianpi Decoction showed an upward trend(P>0.05).The content of propionic acid in the positive control group showed an upward trend(P>0.05).It indicated that the treatment of Hetiao Jianpi Decoction could promote the expression of SCFAs in AAD rats.Conclusions1.Hetiao Jianpi Decoction could improve AAD by repairing intestinal mucosal structural damage,enhancing intestinal mucosal immune function,regulating the abundance of intestinal flora,promoting the expression of SCFAs.2.Repairing intestinal mucosal barrier damage and regulating the balance of intestinal flora were one of the mechanisms of Hetiao Jianpi Decoction in the treatment of AAD. |
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