Influences of Intestinal Microbiota,Motility and TLR4/NF-κB Signaling Pathway by Fecal Microbiota Transplantation in Mice with Refractory Constipation
Objective:To investigate the effect of fecal microbiota transplantation on intestinal microbiota and motility in mice with refractory constipation,whether it was related to Toll like receptor 4(TLR4)/nuclear transcription factor(NF)-κB signal pathway.Methods:60 healthy and clean grade C57BU10 mice,half of males and females,aged 6～8 weeks,with average weight of 22 g.They were randomly divided into three groups,the control group,model group,and fecal microbiota transplantation group,with 20 mice in each group.The model of refractory constipation was replicated by gavage of loperamide hydrochloride(9.38 mg/kg,twice a day,con-secutive for 6 weeks),the number and volume of bowel movements were evaluated to determine the success of model.Fecal microbiota transplantation was performed using 0.2 mL fecal microbiota solution enema of control group mice(twice a day consecutive for 2 weeks).Quantitative detection of microbiota value and B/E value were made by the Japanese Guanggang intestinal microbiota analysis method,intestinal motility was calculated by Evans blue propulsion rate,TLR4,NF-κB,interleukin(IL)-6 and tumor necrosis factor(TNF)-α proteins expression in colon tissues was detected by Western blot.Results:Compared with the control group,the bacterial com-munity values of Enterobacterium,Enterococcus,Bacteroides,and Clostridium in the model group significantly increased,while B/E val-ue decreased;compared with the model group,the bacterial community values of yeast,bifidobacteria,and lactobacilli in the fecal micro-biota transplantation group significantly increased,while B/E values increased(P＜0.05).The Evans blue propulsion rate of the model group was significantly lower than the control group,but it of the fecal microbiota transplantation group was significantly higher than the model group(P＜0.05).What's more,the expression levels of TLR4,NF-κ B.IL-6 and TNF-α proteins in the model group were signifi-cantly higher than the control group,while they were in the fecal microbiota transplantation group were significantly lower than the model group(P＜0.05).Conclusion:Fecal microbiota transplantation could effectively improve the intestinal microbiota and motility in mice with refractory constipation,which may be related to downregulating TLR4/NF-κB signaling pathway activity and inhibition of inflammatory response in intestinal tissues.
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