Probiotic Intervention On Chronic Hepatitis B Patients:A Metagenomic Study

Purpose: Dysbiosis of the intestinal microbiota has been associated with chronic hepatitis B(CHB).We compared the microecological features in different clinical phases of CHB including inactive HBV carrier state,chronic hepatitis,cirrhosis and hepatocellular carcinoma,and identified the potential biomarkers for disease severity.On this basis,three regimes consisting of different combinations of probiotics and prebiotics were used to treat patients with CHB.Liver function,HBV-DNA and HBe Ag clearance were observed.Furthermore,metagenomics was used to characterize the gut flora before and after intervention.Materials and Methods: Part one: Cross-sectional study.We performed a clinical trial of 43 patients including 6 inactive carriers of HBV,17 patients with CHB,14 patients with liver cirrhosis and 6 patients with hepatocellular carcinoma in Shulan(Hangzhou)Hospital from February,2017 to January,2018.43 fecal specimens were tested for metagenomics.Part two: Prospective cohort study.214 patients with CHB were enrolled from 7 hospitals in Zhejiang Province from February,2018 to March,2019.Patients were voluntary to enter the control cohort without intestinal microecological intervention.Other patients were randomly assigned to three groups given different combinations of probiotics and prebiotics: Intervention group 1: probiotics 2g / day for 6 months;Intervention group 2: probiotics 4g / day + prebiotics for 3 months;Intervention group 3: probiotics 4g / day + prebiotics for 6 months.The serum and stool samples of these patients were collected before treatment and 3,6 and 9 months after treatment.The primary endpoints are laboratory indices including HBV-DNA,ALB,ALT,AST before and after intervention.And the secondary endpoints are gut flora before and after intervention assessed by metagenomic sequencing.Results: 1.Fecal samples from inactive carriers of HBV and CHB patients without cirrhosis or hepatocellular carcinoma were enriched with more beneficial flora such as Faecalibacterium prausnitzii,Clostridium leptum,Bifidobacterium and Barnesiella.CHB patients with cirrhosis or hepatocellular carcinoma had higher proportions of Klebsiella,Enterobacter and Escherichia species.In particular,certain viruses seemed to be enriched in the intestine of CHB patients with hepatocellular carcinoma.2.The treatment of probiotics(Bifidobacterium longum,Lactobacillus helveticus)or synbiotics(Bifidobacterium longum,Lactobacillus helveticus,inulin)for 3-6 months had not significantly improved laboratory indices including liver function and HBVDNA in patients with CHB.3.Treatment of synbiotics(Bifidobacterium longum,Lactobacillus helveticus,inulin)for 6 months improved bacterial dysbiosis in patients with CHB featuring abundance of Bifidobacterium and Lactobacillus in feces.Conclusion: Gut microflora dysbiosis happens in different clinical phases of chronic hepatitis B.In a cohort study of patients with CHB,6 months of administration of a symbiotic combination altered the fecal microbiome but did not improve laboratory indices including liver function and HBV-DNA significantly.