The Study On Intestinal Barrier Function And Gut Microbiota In Patients With HBV-associated Liver Disease Based On The "Gut-liver Axis"

Objective:Based on the "gut-liver axis" theory,to explore the changes of intestinal barrier function and gut microbiota in patients with chronic viral hepatitis B（CHB）,posthepatitis B liver cirrhosis（LC）,hepatitis B related acute-on-chronic liver failure（ACLF）and healthy controls.To further clarify the role of "gut-liver axis" in the development of HBVrelated liver diseases.To provide reference for finding new therapeutic targets for chronic hepatitis B,liver cirrhosis and liver failure.Methods:From June 2020 to December 2021,17 patients with CHB,23 patients with LC,and 36 patients with ACLF were enrolled in the department of Infectious Diseases of the First Affiliated Hospital of Soochow University.Ten healthy subjects were included as healthy control group（HC）.General information,clinical indicators such as alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,total bilirubin（T-Bil）,albumin（ALB）,prothrombin time（PT）,platelet（PLT）were collected.To determine whether patients had ascites,gastrointestinal bleeding,hepatic encephalopathy（HE）and other complications.Child-pugh score was also performed in patients with LC or ACLF.Plasma diamine oxidase（DAO）,D-lactic acid（D-LAC）and bacterial endotoxin（ETX）were detected.Fecal genomics of 17 CHB patients,17 LC patients,and 14 ACLF patients were examined using 16SrDNA sequencing.SPSS 25.0 was used to analyze the correlation between intestinal barrier function and relevant clinical indicators in each group.QIIME2 and R3.5.2 were used to analyze the diversity and difference of gut microbiota in each group.Results:Compared with HC group,DAO,D-lactic acid and endotoxin were significantly increased in CHB group,LC group and ACLF group（P<0.05）;Compared with CHB group,DAO,D-lactic acid and endotoxin were significantly increased in LC group and ACLF group（P<0.05）;Compared with LC group,DAO,D-lactic acid and endotoxin were significantly higher in ACLF group（P<0.05）.DAO,D-lactic acid and endotoxin were positively correlated with ALT,AST,T-Bil and PT,but negatively correlated with PLT and ALB（P<0.05）.In LC and ACLF patients,the levels of DAO,D-lactic acid and endotoxin in Child-PughC patients were significantly higher than those in Child-PughB patients,while the levels of DAO,D-lactic acid and endotoxin in Child-PughB patients were significantly higher than those in Child-PughA patients（P<0.05）.DAO,D-lactic acid and endotoxin levels were higher in ascites positive patients and gastrointestinal bleeding patients than in negative control group.DAO and endotoxin levels in patients with hepatic encephalopathy were significantly higher than those in patients without hepatic encephalopathy（P<0.05）.Compared with CHB group,the abundance of Proteobacteria,Prevotella₉ and Enterococcus was significantly increased in LC group（P<0.05）,the abundance of Bifidobacterium decreased significantly（P<0.05）;The abundance of Firmicutes,Proteobacteria,Streptococcus,Veillonella,Escherichia-Shigella and Klebsiella was significant in ACLF group increased（P<0.05）,the abundance of bifidobacteria decreased significantly（P<0.05）;Compared with LC group,the abundance of Firmicutes,Streptococcus and Veroncoccus in ACLF group was significantly increased（P<0.05）,and the abundance of Bacteroidetes,Lactobacillus and Prevotella significantly decreased.（P<0.05）.Conclusion:Intestinal barrier function was impaired in patients with chronic hepatitis B,hepatitis B cirrhosis and hepatitis B-related chronic acute liver failure,and the degree of impairment was closely related to disease severity and complications.The structure and abundance of gut microbiota in patients with chronic hepatitis B,hepatitis B cirrhosis,and hepatitis B-related acute-on-chronic liver failurewere changed,and gut microbiota was associated with the progression of HBV-related liver disease.