Diarrhoea-predominant irritable bowel syndrome distinguishable by 16S rRNA gene phylotype quantification

AIM:To study whether selected bacterial 16S ribosomal RNA(rRNA)gene phylotypes are capable of disting- uishing irritable bowel syndrome(IBS). METHODS:The faecal microbiota of twenty volunteers with IBS,subdivided into eight diarrhoea-predominant (IBS-D),eight constipation-predominant(IBS-C)and four mixed symptom-subtype(IBS-M)IBS patients,and fifteen control subjects,were analysed at three time-points with a set of fourteen quantitative real-timepolymerase chain reaction assays.All assays targeted 16S rRNA gene phylotypes putatively associated with IBS,based on 16S rRNA gene library sequence analysis. The target phylotypes were affiliated with Actinobac-teria,Bacteroidetes and Firmicutes.Eight of the target phylotypes had less than 95%similarity to cultured bacterial species according to their 16S rRNA gene sequence.The data analyses were made with repeated-measures ANCOVA-type modelling of the data and principle component analysis(PCA)with linear mixed-effects models applied to the principal component scores. RESULTS:Bacterial phylotypes Clostridium cocleatum 88%,Clostridium thermosuccinogenes 85%,Coprobacillus catenaformis 91%,Ruminococcus bromii-like, Ruminococcus torques 91%,and R.torques 93%were detected from all samples analysed.A multivariate analysis of the relative quantities of all 14 bacterial 16S rRNA gene phylotypes suggested that the intestinal microbiota of the IBS-D patients differed from other sample groups.The PCA on the first principal component(PC1),explaining 30.36%of the observed variation in the IBS-D patient group,was significantly altered from all other sample groups(IBS-D vs control, P=0.01;IBS-D vs IBS-M,P=0.00;IBS-D vs IBS-C, P=0.05).Significant differences were also observed in the levels of distinct phylotypes using relative values in proportion to the total amount of bacteria.A phy- lotype with 85%similarity to C.thermosuccinogenes was quantified in significantly different quantities among the IBS-D and control subjects(-4.08±0.90 vs -3.33±1.16,P=0.04)and IBS-D and IBS-M subjects (-4.08±0.90 vs-3.08±1.38,P=0.05).Furthermore,a phylotype with 94%similarity to R.torques was more prevalent in IBS-D patients'intestinal micro- biota than in that of control subjects(-2.43±1.49 vs -4.02±1.63,P=0.01).A phylotype with 93%simi- larity to R.torques was associated with control sam- ples when compared with IBS-M(-2.41±0.53 vs -2.92±0.56,P=0.00).Additionally,a R.bromii-like phylotype was associated with IBS-C patients in com- parison to control subjects(-1.61±1.83 vs-3.69± 2.42,P=0.01).All of the above mentioned phylotype specific alterations were independent of the effect of time. CONCLUSION:Significant phylotype level alterationsin the intestinal microbiotas of IBS patients were observed,further emphasizing the possible contribution of the gastrointestinal microbiota in IBS.

Expression of aquaporin 8 in colonic epithelium with diarrhoeapredominant irritable bowel syndrome

Background We analysed and compared the aquaporin 8 （AQP8） expression in ascending and descending colon mucosa between patients with diarrhoea-predominant irritable bowel syndrome （D-IBS） and healthy volunteers, in order to study the relationship between the clinical feature of IBS, the expression of AQP8 and the pathological mechanism of D-IBS. Methods Specimens were taken from the proximal ascending colon or distal descending colon of D-IBS patients （n=-26） and healthy volunteers （n=30）, and AQP8 mRNA expression of each specimen was determined by fluorescent quantitative reverse transcription polymerase chain reaction （FQ-RT-PCR）. In patients with D-IBS, the relationship was analysed between AQP8 expression in both ascending and descending colons and clinical features including gender, age of onset, duration of illness, frequency of defecation, and stool characteristics. Results Although AQP8 was present in the epithelium of the ascending and descending colons in healthy persons and D-IBS patients, the AQP8 level of the D-IBS patients was significantly lower than that of the healthy persons （P〈0.01 in the ascending colon, P〈0.05 in the descending colon）. AQP8 expression was not correlated with the age of patients with D-IBS （P〉0.05 both in the ascending and descending colons） or the age at the onset （P〉0.05 both in the ascending and descending colons）, but closely with the duration of illness （P〈0.05 in the ascending colon, P〈0.01 in the descending colon）, frequency of defecation （P〈0.01 in the ascending colon, P〈0.05 in the descending colon） and stool characteristics （P〈0.01 in the ascending colon, P〉0.05 in the descending colon）. Conclusions The decreased AQP8 expression in D-IBS patients indicates that dysfunction of colonic absorption may cause reduced water absorption, loose stool and diarrhoea. The expression of AQP8 may be related to D-IBS.