Study On The Imbalance Of Biliary Flora In Patients With Hepatolithiasis

Background: To study the characteristics of biliary microflora and its relationship with the occurrence and development of hepatolithiasis.Methods: Forty patients with hepatolithiasis who needed partial hepatectomy were recruited as the experimental group and 10 patients with hepatic space occupying lesions as the control group.The relevant clinical data were recorded,including infection index,liver function index,histopathology and so on.Bile samples were collected during the operation and preserved at-80 ℃.The DNA of bile sample flora was extracted,and the hypervariable region V4 of DNA was amplified by PCR with specific primers,and the gene was sequenced.The sequencing results were clustered by operational taxon(OTU).The representative sequences of OTUs were compared with SILVA database,and species annotations were made,and then biliary flora abundance analysis,α diversity analysis,β diversity analysis and other aspects were studied.Results: A total of 44 bile samples from participants were tested for bacteria,including 40 in the experimental group and 4 in the control group.There were differences in direct bilirubin(24.82 ±40.57 vs 4.48 ±3.67,P = 0.003),indirect bilirubin(22.54 ±24.27 vs 11.33 ±5.10,P = 0.010)and γ-glutamyl transpeptidase(281.37 ±280.66 vs 77.85 ±76.79,P < 0.0001)between the experimental group and the control group.There was a significant difference in bile flora spectrum between the experimental group and the control group.At the level of bacterial phylum classification,the highest abundance in bile of the experimental group and the control group were Proteus(52.4% VS 40.2%)and Streptomyces(42.1%VS 29.6%).At the classification level of bacteria,Enterococcus was the most abundant in bile of the experimental group(33.77%),and Bifidobacterium was the highest in the bile of the control group(6.0%).The diversity of microbial communities in bile of patients with hepatolithiasis decreased significantly Shannon(P = 0.004),Observed species(P=0.001),PD-whole-tree(P= 0.001).In addition,there were intra-group differences among the samples in the experimental group,but the intra-group difference in the experimental group was smaller than that in the control group.Enterococcus,unclassified Enterobacter and Lachnoclostridium had significant differences between the two groups,which were 24.68%,12.35% and 8.80%,respectively.There was a significant difference in phyllotaxonomy between the experimental group and the control group(P = 0.007),while at the taxonomic level,it was mainly Enterococcus(P < 0.001)and unclassified＿Enterobacteriaceae(P = 0.003).Conclusion:The biliary tract of patients with hepatolithiasis contains a large number of bacteria and dysbacteriosis.The imbalance of biliary flora may be related to the pathogenesis of hepatobiliary calculi.Bacteria are also found in bile in healthy bile ducts.