Effect Of Three Food Extracts On The Regulation Of Intestinal Function Of An Antibiotic Induced Constipation Mouse Model

The constipation is found to be induced by multiple of factors such as the disorder of intestinal microbiota,inflammation,malfunction of intestinal motility and the deficiency of intestinal nutrition.The imbalance of intestinal microbiota caused by lifestyle,dietary structure and antibiotics,etc can leads to constipation.Patients with constipation have intestinal microbiota disorder.The prevalence of constipation in the population has reached27% and it presents an upward trend,according to the data from surveys.Long term constipation can cause many complications,which leads to anorectal diseases,gastrointestinal dysfunction etc.However,the treatment of constipation at home and abroad has shown many disadvantages,such as strong drug dependence,side effects and chronic toxicity.Therefore,it is very important to find a way to treat constipation with remarkable,comprehensive,green and safe functions.Natural food ingredients have become a new direction for research due to the characteristics of less harm to human health,a variety of function to living organisms and a good therapeutic function for diseases.In this study,several food extracts were selected to explore the regulatory effects on intestinal function,low-grade inflammation,intestinal microbiota and other functions,and the alleviation to constipation.The correlation of the alleviation of constipation between the improvement of intestinal function by food extracts and improvement of intestinal microbiota by food extracts was explored by applying ELISA kit and 16 S r DNA sequencing technology.The correlation the change of intestinal microbiota structure and constipation was also explored.This study provides new ideas and theoretical data guidance for the further development of new constipation products.The research results and details are as follows:1.The establishment of a model of intestinal microbiota disordered mouse constipation induced by antibioticsThe model of constipation with intestinal microbiota disorder was established by the intragastric administration of mixture suspension of clarithromycin,cephalexin,and amoxicillin.The procedure includes intragastric administration of suspension of 62.5mg/m L for 4 d and 125 mg/m L for 3 d.The results showed difficulty in defecation,stool shape was dry and unable to granulate,and small intestine peristalsis function were weakened.In the constipation model group,the excretion volume within 1 hour and dailyaverage excretion volume decreased significantly(P<0.01).The feed consumption rate decreased significantly(P<0.01).The fecal moisture content decreased significantly(P<0.01),and the duration of the first black stool was significantly extended(P<0.01)Besides,the intestinal propulsion rate was significantly reduced(P<0.01).The measurement of intestinal microbiota in mice showed that the diversity of intestinal microbiota in the constipation model group was significantly reduced,and the species abundance was lower than that in the blank group.In terms of species composition at the phylum level,the dominant phylum of the constipation model group is Bacteroidetes,and the phylum of Verrucomicrobia was absent.In terms of species composition at family level,the constipation model group was mainly composed of Bacteroidaceae,O-Clostridiales,Lachnospiraceae,Enterobacteriaceae,and the relative abundance decreases.The constipation model group has a significantly lower level of species than the blank group,which is mainly composed of Bacteroides,f-Lachnospiraceae,f-Peptostreptococcaceae,f-Enterobacteriaceae,Trabulsiella,and the abundance of O-Clostridiales,f-Enterobacteriaceae,f-S24-7 was significantly reduced.This indicates that antibiotics have caused the disorder of intestinal microbiota in mice,and the murine model of constipation with intestinal microbiota disorder was successfully established.2.Regulation effect of three food extracts on intestinal function of antibiotic-induced constipation mouse modelThe results of the study showed that after 7 days of intervention with three food extracts,compared with before intervention,the small intestine advancement rate increased,the duration of the first black stool was shortened.And the mouse fecal moisture content,the first black stool duration,and the small bowel advancement rate were significantly increased(P<0.01).This indicated that food extracts significantly improve the quality of feces,the function of small intestine peristalsis and speed up the transmission frequency of small intestine.The expression levels of several inflammatory factors showed that the expression of three inflammatory factors IL-1β,IL-6,and TNF-α in serum were significantly reduced(P<0.01).The concentration of LPS in mouse serum was significantly reduced(P<0.01)and the expression level of the anti-inflammatory factor IL-10 was significantly increased(P<0.01).The IL-1β concentration in the serum of mice in the low-dose mussel group was significantly reduced,the IL-6 in the serum of mice in the high-dose shrimp head group was significantly reduced,the concentration of TNF-α in the high-dose shrimp head group was significantly reduced,and the concentration of LPS in the low-dose group of compound formula was significantly reduced when compared to blank group.3.Regulation effect of three food extracts on intestinal microbiota in mouse model of constipation induced by antibioticThe result of 16 S rDNA sequencing showed that three food extracts changed the structure of mice intestinal microbiota and also increased the abundance and diversity of community.Compared with the constipation model group,the species uniformity of intervention group flora of intervention group was high and the number of OTU was high.Result of the abundance composition at different species classification levels of constipation group showed that at the phylum level,Bacteroidetes from constipation model group was the dominant bacteria,the beneficial bacteria Verrucomicrobia was missing,Proteobacteria from intervention group was the dominant bacteria,Proteobacteria,Verrucomicrobia increased significantly when compared to blank group.At the family level,Enterobacteriaceae was the dominant intestinal bacterium in the intervention group,and Bacteroidaceae was the dominant intestinal bacteria constipation group.The abundance of beneficial bacteria Verrucomicrobiaceae,Rikenellaceae,and Enterobacteriaceae increased.At the genus level,the beneficial bacteria in the intervention group were Akkermansia,Trabulsiella,f-Rikenellaceae,Blautia,Enterobacteriaceae;the dominant bacteria in constipation group were Bacteroides.The results of Akkermansia,Enterobacteriaceae,Bacteroides and typical inflammatory factor TNF-α,anti-inflammatory factor IL-10,endotoxin(LPS)expression levels Correl index indicate that Akkermansia,Enterobacteriaceae and inflammatory factor TNF-α have a negative correlation.It also demonstrated a strong correlation between Enterobacteriaceae and TNF-α.Akkermansia,Enterobacteriaceae have a positive correlation with IL-10 while Akkermansia and IL-10 have a strong correlation.Bacteroide has a strong positive correlation with LPS.This indicates that with the increase in the abundance of Akkermansia and Enterobacteriaceae,the two bacteria with the ability to suppress inflammation,and the decrease in the abundance of Bacteroides,leads to the significant decrease inflammatory level in mice.In summary,three food extracts increase the abundance and diversity of intestinal microbiota,the relative abundance of beneficial bacteria in intestine and inhibit the growth of harmful bacteria in the intestine.Thus,food extracts improve inflammation,and maintain the steady state of the intestinal microbiota structure.Among which the low-dose mussel,low-fructose oligosaccharide,and high-dose banana groups adjusted the intestinal microbiota significantly.