A comparative study of intestinal flora between hepatitis B cirrhosis patients with or without ascites

Objective To investigate the difference in intestinal flora between hepatitis B liver cirrhosis( HBLC) patients with or without ascites. Methods A total of 57 patients with HBLC who visited Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from October to December 2016 were enrolled,among whom 30 had no ascites( HBLC-WOA group) and 27 had ascites without spontaneous peritonitis( SBP)( HBLC-WA group),and 28 healthy volunteers were enrolled as healthy controls( HC group). Intestinal flora was compared between the two groups of HBLC patients using 16 S rRNA sequencing. The Wilcoxon rank sum test was used for comparison of categorical data between two groups,and the Mann-Whitney U test was used for comparison of continuous data between two groups;the Kruskal-Wallis H test was used for comparison of continuous variables between more than two groups. Multiple hypothesis tests were used for comparison of relative abundance between species and was adjusted by Benjamini and Hochberg false discovery rate( fdr),and Pfdr< 0. 05 was considered statistically significant. The Spearman rank correlation test was used for correlation analysis. An analysis of similarity( ANOSIM) and a non-parametric multivariate analysis of variance( Adonis) were used for comparison between groups under different conditions. Results The abundance of fecal microbiota gradually decreased with the appearance of ascites in HBLC patients( P = 0. 042).There were significant differences between the HBLC-WOA group and the HC group( ANOSIM: R = 0. 159,P = 0. 001; Adonis: R2=0. 067,P = 0. 001) and between the HBLC-WA group and the HC group( ANOSIM: R = 0. 323,P = 0. 001; Adonis: R2= 0. 107,P =0. 001). At the genus level,compared with the HC group,the HBLC-WA group had significant reductions in the abundance of Subdoligranulum and Pseudobutyrivibrio( P < 0. 01 and P < 0. 001) and significant increases in the abundance of Enterobacter,Escherichia,and Veillonella( P < 0. 05,P < 0. 001,and P < 0. 01). Escherichia and Veillonella were positively correlated with Child-Turcotte-Pugh( CTP) score,prothrombin time,and international normalized ratio and were negatively correlated with serum albumin( Alb) level( all P <0. 05). Pseudobutyrivibrio,norank ＿f ＿ Lachnospiraceae,unclassified ＿ f ＿ Lachnospiraceae,and Blautia were positively correlated with Alb level and were negatively correlated with CTP score and C-reactive protein level( all P < 0. 05). The KEGG pathway analysis showed that with the appearance of ascites,there were gradual increases in the abundance of the pathways associated with transcription-related proteins,alpha-linolenic acid metabolism,Staphylococcus aureus infection,bacterial invasion of epithelial cells,and bile secretion( all Pfdr< 0. 05),as well as a gradual reduction in the abundance of the pathway associated with the biosynthesis of flavonoids( Pfdr< 0. 05). The pathway associated with bacterial invasion of epithelial cells was positively correlated with the abundance of Escherichia( P < 0. 001),and the abundance of Enterobacter was positively correlated with the pathway of bile secretion( P < 0. 001). Conclusion Intestinal flora disturbance is observed in HBLC-WA patients,featuring the reductions in the abundance of Subdoligranulum and Pseudobutyrivibrio belonging to Firmicutes and the increases in the abundance of Enterobacter and Escherichia belonging to Proteobacteria. Enterobacter may be involved in the pathway of bile secretion,and Escherichia may be involved in the pathway associated with bacterial invasion of epithelial cells. It is suggested that regulation of intestinal flora,as well as the prophylactic treatment of SBP,should be considered for HBLC-WA patients without SBP.