The Gut Flora Structure And Function Of The Patient's Are Affected By Antibiotics And Probiotics Therapy

The gut flora of the patient has a profound influence on the microbiological environment of the hospital.Medical antibiotics are one of the main sources of antibiotics in the environment.Due to the unique metabolic characteristics of antibiotics,the residues of antibiotics in environmental media,including water bodies and soil,have resulted in a wide and far-reaching impact on different organisms and ecosystems.With the rapid development of social economy,the incidence rate of liver toxic damage,toxic hepatitis and liver cirrhosis with mild and micro hepatic encephalopathy has increased due to the changes of environmental pollution toxins,people's living habits and lifestyles.In this study,the structure and function of intestinal microflora of patients with liver cirrhosis with mild micro-hepatic encephalopathy were studied under the influence of antibiotics and probiotics to reveal the changes of intestinal microflora before and after the treatment of mild micro-hepatic encephalopathy with antibiotics and probiotics.At the same time,it will be of great significance for us to explore the therapeutic mechanism of hepatic encephalopathy.In this study,rifaximin combined with probiotics was used to treat patients with MHE,and the gut flora of patients with minimal hepatic encephalopathy before and after treatment was analyzed by macrogenomics.To reveal the changes of gut flora structure and function in patients with MHE treated with rifaximin plus probiotics.The clinical indexes of serum inflammatory factors,blood ammonia,liver function,coagulation function and neuropsychology before and after treatment were compared and analyzed.The clinical efficacy of rifaximin combined with probiotics in the treatment of MHE and its mechanism were discussed.21 patients with mmimal hepatic encephalopathy of liver cirrhosis were randomly divided into two groups: rifaximin combined with probiotics therapy group(n=9)and rifaximin treatment group(n=12)as control group.Fecal samples were collected from patients with minimal hepatic encephalopathy before and after treatment.16 s r DNA of faecal flora was extracted.High throughput sequencing was carried out based on Illumina Hi Seq sequencer.The results were analyzed by bioinformatics.The serum inflammatory factors,blood ammonia,liver function,coagulation function and neuropsychological parameters were measured simultaneously before and after treatment.And carries on the multivariate statistical analysis to the data.The main research results are as follows:By bioinformatics analysis,a total of 1625975 high quality reading sequences were obtained from 42 samples.The average number of reading sequences per sample was 38714±812(SEM).Using 97% sequence similarity as the classification criterion at the species level,a total of 1684 OTUs,an average of 317±29(SEM)OTUs per sample were identified.The 1684 OTUs of the whole data set belong to 243 genera of 29 gates.The sequence coverage of the whole sequence data set is 99.69% ±0.03%(SEM).In order to reveal the influence on intestinal microbial structure of MHE patients before and after treatment,we analyzed the level of intestinal flora classification,genus and species.(1)Intestinal flora classification level analysis,through the sequence analysis and calculation of index of diversity of species classification and,before and after treatment in patients with MHE intestinal flora Alpha diversity comparison and analysis of statistics,intestinal flora of clustering analysis,statistical analysis results show that the Alpha diversity,Shannon diversity index analysis,Good 's coverage of sequencing coverage index analysis,before and after two treatments,MHE patients with intestinal flora classification level were no significant difference.(2)At the level of gut bacterial species genus,the influence on the intestinal microbial structure of MHE patients before and after treatment.From the analysis of the level of intestinal flora and the method of PAM clustering analysis,three types of intestinal flora were identified: Escherichia(cluster-E),Streptococcus(cluster-S)and Bacteroides(cluster-B).The results showed that there was no significant difference in the level of intestinal flora in MHE patients before and after the treatment of rifaximin alone and rifaximin combined with probiotics.(3)At the level of gut bacterial species,based on the distance measure Unifrac system development(including the unweighted Unifrac and weighted Unifrac),through the principal coordinate analysis(principal coordinates analysis,PCo A)and displacement Multivariate Variance test analysis(PERMANOVA,Permutational Multivariate analysis of Variance)are two kinds of therapy before and after treatment,variation degree of intestinal micro organism structure in patients with MHE.The results showed that there was no significant difference in the level of intestinal flora in MHE patients before and after the treatment of rifaximin alone and the combination of rifaximin and probiotics.In order to reveal the MHE patients before and after the treatment effect of intestinal flora function,we use PICRUSt method,from the level of metabolic pathways,KO level(KO,KEGG Orthology,Kyoto encyclopedia orthologous genes and genome genetic classification system)are analyzed using two methods of treatment for the degree of variation gut microbial metabolic function in patients with MHE.The intestinal flora function in patients with MHE treated by rifaximin alone showed no significant changes in metabolic pathway level and KO level before and after treatment.On the contrary,The intestinal flora function in patients with MHE treated by rifaximin combined with probiotics showed significant changes in metabolic pathway level and KO level before and after treatment.The patients with MHE were treated by rifaximin and rifaximin combined with probiotics,neuropsychological examination:digital connection test A(NCT-A),digital symbol test(DST),were significantly improved before and after treatment(P< 0.05).The treatment of rifaximin and the combination of rifaximin with probiotics can significantly reduce the blood ammonia concentration in patients with MHE.The serum inflammatory index of MHE:IL-2,IL-4,IL-6,IL-10,TNF-?,CRP was significantly decreased before and after treatment(P<0.05).The liver function and coagulation function were improved after the treatment(P<0.05).RDA(redundancy analysis)was used to analyze the correlation between intestinal flora and clinical data in different treatment schemes.The level of serum IL-2,IL-6,TNF-? in patients with MHE,treated with rifaximine was significantly correlated with intestinal flora before and after treatment.In combination therapy,IL-2 levels were significantly correlated with intestinal flora before and after treatment.In conclusion,this study showed that the combination of rifaximin and probiotics did not affect the structure of intestinal flora in patients with MHE,but significantly changed the function of intestinal flora in MHE patients.Rifaximin had no effect on the structure and function of gut flora in MHE patients.Rifaximin combined with probiotics and rifaximin alone has good clinical efficacy in patients with MHE.The mechanism of the effect of rifaximin combined with probiotics on MHE may be one of the mechanisms of clinical efficacy.Reducing inflammation in patients with MHE is one of the mechanisms of two therapies for MHE.The mechanism of rifaximin plus probiotics in reducing blood ammonia concentration in patients with MHE may be due to the synergistic effect of antibiotics and probiotics on the cultivation of potentially beneficial gut environment.