The Study Of Gut Microbiome In Chronic Functional Constipation And In Synbiotics Treatment

ObjectiveTo observe the structure of intestinal microbiome in patients with chronic functional constipation,and the use of multiple omics research in synbiotics treatment of chronic functional constipation,observing the changement in clinical effect,the intestinal microbiome and intestinal metabolic environment.Methods（1）in department of microbial treatment of Shanghai tenth people’s hospital,patients who diagnosed with chronic functional constipation and matched the include and exclude standards can be included in the research,and then register the related clinical data of patient,and collect the patient’s feces.informed consent about joining the synbiotics treatment should be signed before the research according to the ethical requirements,synbiotics preparations containing 1 x 10¹⁰CFU/g probiotics,soluted with warm water after each meal in 30 min and used last for 1 month,then record bowel habit,complications of constipation and collect the feces of patients again after synbiotics treatment.people who volunteer to take part in for last 3 months are also should be recorded the data of bowel habit,complications of constipation and collected the feces.at the same time,the similar required data and fecal samples are collected from healthy volunteer as a control group.（2）the multiple-omics analysis were taken to the feces collected,firstly,all samples for were taken by 16S-rRNA sequencing,（extracted the genomic DNA from feces,sequenced for V4-V5 area）,on the basis of the results of 16S sequencing,metagenome sequencing were taken on selected fecal samples to further identificate of bacteria（through the shotgun method）.Metabonomics detection is taken on all samples at the same time,to identify small molecular metabolism in feces with the method of non-targeting.Results（1）53 cases of patients were intaked in chronic functional constipation group,31 cases of men and 22 cases of women,defecation frequency per week was 2.22±0.09 times on average,clinical history of constipation was 105.60±35.95 months.After synbiotics treatment for 1 month,there were still 36 people who willing to accept follow-up and return fecal samples to hospital,after treatment after 3 months,there were also 15people,the control group were taken by 53 healthy community volunteer.29 cases of male and 24 case of female,defecation frequency per week was 8.06±0.34 times on average,and with no clinical history of constipation.the defecation frequency per week was increased after the synbiotics treatment,and the occurrence rate of abdominal distension,loss of appetite,laxative dependence were decreased.the difference was statistically significant（P<0.05）.no adverse reactions occured in the process of treatment.（2）according to the results of 16S-rRNA sequencing analysis,genus of Prevotella₉had a lower abundance while genus of Escherichia-Shigella had a higher abundance in constipation group,after synbiotics treatment inconstipation group,the abundance of Prevotella₉ increased and which of Escherichia-Shigella decreased,according to the metagenomic data analysis,the abundance of Bifidobacterium_dentium was enriched in healthy group,however,another two species of Bifidobacterium（Bifidobacterium_adolescentis and Bifidobacterium_animalis）were enriched in constipation group.however.the abundance of two species of Lactobacillus（Lactobacillus_oris and Lactobacillus_reuteri）were enriched in healthy people.after the synbioticstreatment,the abundance species of Bifidobacterium_animalis and Pseudomonas_aeruginosa were increased.From the results of the untargeted metabolomic analysis,metabolites like acetic acid were enriched in healthy group,and which was increased after treatment.Conclusioncompared with the composition of intestinal microbiome in healthy group.the intestinal microbiome structure had changed significantly in chronic functional constipation group,not only in aspect of the microbiome changed but also in the related metabolites changed.After with the synbiotics treatment in constipated patients,it not only achieved the improvement in clinical effect,but also can carry on optimization to the intestinal microbiome.This study with the perspective of clinical trials found that microbial treatment has a positive meaning and application prospects...