Alteration Of Gut Microbiome With Chronic Liver Diseases And Application For Diagnosis Of Liver Cancer

ObjectiveTo analyze the characteristics of gut dominant microbiota comparatively of liver cirrhosis,liver cancer and autoimmune liver disease and its correlation with clinical indexes.To screen the key gut microbiome index to distinguish between liver cancer and other chronic liver diseases.MethodsA total of 117 patients and 30 healthy controls were enrolled from January 2014 to February 2017 at the First Affiliated Hospital of Zhejiang University Medical College and Hangzhou Shulan Hospital.According to the clinical diagnosis,the patients were divided into three groups:liver cirrhosis(including hepatitis B cirrhosis and alcoholic cirrhosis),liver cancer and autoimmune liver disease(including primary biliary cirrhosis and autoimmune hepatitis).We analyze the clinical manifestation,laboratory index,Child-Pugh score,MELD score,characteristics and differences of gut microbiota of patients,and the correlation of clinical indicators and gut dominant microbiota.The single factor analysis of variance(ANOVA)was used to analyze the difference of gut dominant microbiota among different groups.The correlations was analyzed by Pearson correlation coefficient.Logistic regression was used to analyze and filter the specific gut bacteria of liver cancer.The index model was established by ROC curve fitting Test,and calculate its sensitivity and specificity.ResultsIn this study,117 patients with chronic liver disease were enrolled in this study.The patients were mainly in the elderly population,the average age 55.4 ± 12 years old,male/female=1.39:1.Liver cirrhosis and liver cancer patients with male-based,the proportion was 79.5%and 81.6%,respectively,but the autoimmune liver disease in patients with female-based(94.3%).There were very significant differences in Child-Pugh score and MELD score between liver cancer and liver cirrhosis(P<0.001).Liver cancer and cirrhosis patients with systemic symptoms and gastrointestinal discomfort as the main clinical manifestations,and more than 50%patients with portal hypertension manifestations.The incidence of complications in cirrhosis group was higher than that in liver cancer group.Liver cancer,liver cirrhosis,autoimmune liver disease and healthy control group showed at the Faecalibacterium prausnitzii,Enterococcus,Bacteroides,Bifidobacterium,Lactobacillus,Clostridium leptum,Eubacterium rectale,Enterobacteriaceae and B/E Values were very significantly different(P<0.00).Compared with the healthy control group,Bifidobacterium and B/E were significantly decreased in the three case groups,and the Enterobacteriaceae were significantly increased.Clostridium leptumin cirrhosis patients was significantly reduced.Compared with the other three groups,the content of Enterococcus in the autoimmune liver disease group was significantly decreased and the number of Bacteroides was significantly increased.Compared with the other three groups,with anti-inflammatory effect of F.prausnitzii in the gut is relatively small and E.rectale more in patients with liver cancer.The results of correlation analysis between intestinal flora and laboratory index showed that in cirrhosis group,both F.prausnitzii and E.rectale were positively correlated with alanine aminotransferase(ALT)and aspartate aminotransferase(AST),and was negatively correlated with CRP.The correlations of Bateroides and Clostridium leptum with white blood cell count(WBC),neutrophil absolute value(NEUT#),CRP,and correlations between Bacteroides and Child-Pugh score,Clostridium butyricum and CRP were negatively,and was positively between Enterococcus and CRP.In liver cancer group,correrlations of F.Prausnitzii with(NEUT%)and fibrinogen(FBG),E.rectale with NEUT%,high sensitivity C-reactive protein(CRP),FBG,prothrombin time(PT)and Child-Pugh and MELD scores,Enterococcus with platelet(PLT),Bacteroides with FBG,were negatively.While E.rectale was positively correlated with ALB.and the correlation between the indicators was moderately correlated in liver cancer patients.Using Logistic regression in univariate analyses to filter the the specific gut bacteria of liver cancer.The index is the ratio of E.rectale to F.Prausnitzii(1gEr/1gFp).Both the 1gEr/1gFp and the serum alpha-fetoprotein(AFP)can distinguish between liver cancer and without liver cancer patients.Moreover,the 1gEr/1gFp can identify liver cancer patients with AFP values in the normal range and the cirrhosis cases with a higher AFP(P<0.01).The results of binary logistic regression analysis of AFP and 1gEr/1gFp showed that the 1gEr/1gFp(P<0.05)was more valuable for the diagnosis of liver than AFP(P?0.05).The results of ROC curve fitting were performed between the two indexes and the combination of the two indicators:For liver cancer and cirrhosis,the AUC of combine with 1gEr/1gFp and AFP was largest(AUC=0.786),what means the combine can diagnosis of liver cancer best effect,and the 1gEr/1gFp(AUC=0.748)is better than AFP(AUC=0.664),and the specificity of 1gEr/1gFp is higher(97.7%).In the two groups of liver cancer and autoimmune liver disease,the combined detection(AUC=0.949)of the two is the best for the diagnosis of liver cancer;And its sensitivity and specificity were 81.6%and 100%respectively.For liver cancer and liver cirrhosis with high AFP,liver cancer with low AFP and cirrhosis,the AUC of 1gEr/1gFp is greater than 0.8,so the 1gEr/1gFp has a certain accuracy to diagnose the liver cancer.And 1gEr/1gFp for the distinction between liver cancer and high AFP liver cirrhosis specificity was-93.3%,while the distinction between low AFP liver cancer and cirrhosis of the sensitivity is 76.5%.For liver cancer and non-liver cancer patients,the 1gEr/1gFp and AFP combined detection(AUC=0.804)have the best effect to diagnose the liver cancer,followed by 1gEr/1gFp(AUC=0.752).The specificity of 1gEr/1gFp and combined detection of liver cancer diagnosis is up to 98.4%.ConclusionsLiver cancer,liver cirrhosis and autoimmune liver disease in patients with gut dominant microbiota structure has undergone significant changes,the beneficial bacteria Bifidobacterium significantly reduced,and the Enterobacteriaceae significantly increased.In the development process of liver cirrhosis to liver cancer,the gut microbiome may be suffered some changes.In liver cancer group,with anti-inflammatory effect of F.prausnitzii in the gut is relatively small and E.rectale more in patients with liver cancer.In Liver cancer and cirrhosis patients,the relations of gut microbiota with inflammation liver function and coagulation function is highly correlated.For the value of diagnosis of liver cancer,the gut microbiome marker 1gEr/1gFp is more than that AFP.The 1gEr/1gFp can distinguish the liver cancer patients with normal AFP values and the cirrhosis patients with a higher AFP.Overall,the specificity of 1gEr/1gFp and combined detection of liver cancer diagnosis is better than AFP,and the combined detection of 1gEr/1gFp and AFP have the best effect to diagnose the liver cancer.