Study On The Characteristics Of Gut Microbiome In Patients With Chronic Prostatitis/Chronic Pelvic Pain Syndrome

BackgroundChronic prostatitis/chronic pelvic pain syndrome(CP/CPPS)is the most common urinary system disease faced by men under the age of 50.In 1995,the National Institutes of Health(NIH)proposed a new classification standard to play a guiding role in the research and clinical practice of prostatitis.Prostatitis is divided into four different types by the NIH classification standard.The most common type is CP/CPPS,which is also known as chronic nonbacterial prostatitis.CP/CPPS is a group of common urological diseases caused by multiple etiologies and with various clinical manifestations.The pathogenesis of chronic prostatitis/chronic pelvic pain syndrome has not been elucidated.It may be caused by a variety of etiologies,including pathogen infection,urinary dysfunction,neuroendocrine factors,pelvic venous congestion,and abnormal immune response.Its clinical manifestations are mainly lower urinary tract dysfunction,including urination symptoms,pain symptoms and mental symptomsIn recent years,culture-independent bacteria identification techniques have allowed us to identify the microbial composition in intestinal/urinary tract specimens,and can link the characteristics of intestinal/urinary tract microbiome to specific diseases Based on this,there have been many studies in recent years to explore the relationship between chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS)and the microbiome of the urogenital tract,but the results have not provided new insights for the its diagnosis and treatment Ideas.At the same time,in recent years,there have been many positive discoveries in the field of gut microbiome.At present,between patients with digestive system,metabolic diseases,nervous system diseases,cardiovascular system diseases,respiratory system diseases and other diseases,and the control group,there are differences in the composition and function of the gut microbiome have been confirmed.Based on these latest findings,manifestations of autoimmune disorders and hypersensitivity of pain that may be involved in the pathogenesis of CP/CPPS have been confirmed to be related to the disturbance of gut microbiome.Therefore,we suspect that the imbalance of gut microbiome may be related to the occurrence and development of CP/CPPS.At present,there are few reports on the phenotype,mechanism and function of gut microbiome in CP/CPPS,so we carried out this clinical study.In this study,we used 16srDNA sequencing technology and related bioinformatics analysis and statistical analysis to explore the structural differences between the gut microbiome of CP/CPPS patients and asymptomatic healthy volunteers,and tried to discover the characteristics of gut microbiome composition of CP/CPPS patients,To provide a new direction for the understanding of the pathogenesis of the disease and the potential diagnosis and treatment direction.MethodsFrom March 2018 to February 2019,the recruitment of multi-center research objects was carried out.CP/CPPS patients were recruited from patients who were treated in the urology clinic of Qilu Hospital of Shandong University,Shandong Provincial Hospital and Qianfoshan Hospital of Shandong Province.A healthy control group was recruited from the healthy family members of patients from Shandong University Qilu Hospital Ward and student volunteers from Shandong University.The main exclusion criteria for this study were the history of intravenous or oral antibiotic application in the last 3 months.All subjects who met the inclusion and exclusion criteria signed an informed consent.Conduct background information questionnaire survey on the research subjects who meet the inclusion and exclusion criteria.We use NIH-CPSI scores,urine culture,urine routine and prostatitis routine test CP/CPPS patients for screening,symptom severity assessment and clinical classification.Stool samples from the CP/CPPS group and the control group were collected for bacterial-specific 16S rDNA gene sequencing.Through bioinformatics analysis and statistical analysis,analyze the general data of the research objects,and assess the diversity,species composition,and species differences between the two groups of gut microbiome.We analyzed the correlation between the differential bacteria and the clinical manifestations of CP/CPPSResultsa General patient information.There were 41 people in the CP/CPPS group with an average age of 26.27 years(±7.34)and 42 healthy controls,with an average age of 25.56(±7.70).There was no statistically significant difference between the two groups(P>0.05)b In this study,high-throughput sequencing was performed on the v3-v4 region of gut microbiome 16SrDNA of CP/CPPS group and healthy control group,and a total of 4656152 effective sequences were obtained;the rarefaction curve reflects that the amount of sequencing data is reasonable and sufficient.c There was no significant difference in the gut microbiome Alpha diversity between CP/CPPS group and healthy control group.d The composition of gut microbiome in CP/CPPS group and healthy control group are different.The number of overlapping genera in the gut microbiome of the two groups was 337,that of the CP/CPPS group was 39,and that of the healthy control group was 48.In terms of the difference in species composition and abundance of gut microbiome between the two groups,the community bar graph shows that at the genus level,the mean value of the abundance of each genus is different.e There is a difference in the beta diversity of gut microbiome between the CP/CPPS group and the healthy control group.PCoA analysis showed that there were differences in the coordinate distribution of gut microbiome between the two groups,suggesting that there was a difference in Beta diversity between the two groups.f Significance test of differences between groups at single taxonomic level.Compared with the healthy control group,the gut microbiome structure in the CP/CPPS group had statistically significant differences in abundance:at the classification level of the Phylum,the proportion of Proteobacteria was increased(P=0.007);at the classification level of the Class,the Gammaproteobacteria proportion increased(P=0.018),Betaproteobacteria proportion decreased(P=0.029);at the classification level of the Genus,the proportion of Lachnoclostridium increased(P=0.018),while Dorea(P=0.0354),Eubacterium hallii(P=0.0094),Alistipes(P=0.0005),and Dialister(P=0.0257).g Discriminant analysis of Lefse multi-level species differences at the multi-taxonomic level.The predominant floras in the CP/CPPS group include Gammaproteobacteria,Proteobacteria,Lachnoclostridium.And the predominant floras in healthy control group include Alistipes,Rikenellaceae,Dialister,Dorea,Betaproteobacteria,Burkholderiales and Eubacterium_hallii_group.These differences are statistically significant.h Spearman correlation analysis found that the abundance of Lachnoclostridium was moderately positively correlated with the NIH-CPSI score(rho=0.40,P=0.009),indicating that the abundance of Clostridium is moderately correlated with the clinical symptoms of CP/CPPS patients.Conclusiona Most of the bacterial species in the gut microbiome of the CP/CPPS group and the healthy control group are shared by the two groups,and a few of them are unique to the two groupsb There is a significant difference in gut microbiome between CP/CPPS group and healthy control group.The composition of the gut microbiome of the two groups is different,which shows that there are significant differences in the abundance of multiple bacteria at different taxonomic levels.c Among them,Lachnoclostridium may play a role in the pathogenesis of CP/CPPS.