Title Page

Contents

Abstract English 10

Ⅰ. Introduction 12

Ⅱ. Materials and Methods 14

1. Human Study 14

2. Stool sample collection and analysis 15

3. Bacteria culture (Akkermansia muciniphila) 16

4. Animal model by DDC treatment 17

5. Intestinal permeability analysis 18

6. Endotoxin assay & Cytokine Measurement 18

7. Realtime-Polymerase Chain Reaction analysis 19

8. Quant-Seq microarray 19

9. Immunofluorescence 20

10. Serotonin ELISA 21

11. Statistical analysis 21

Ⅲ. Results 22

1. Depend on severity of Cirrhosis patient the composition of human gut microbiome alterations. 22

2. Akkermansia muciniphilla can ameliorate DDC-induced liver fibrosis 26

3. A.muciniphilla has better effect on ameliorate liver fibrosis in Humanized mouse model. 28

4. A.muciniphilla modulate the gut bacteria composition and diversity in DDC diet model. 30

5. Involvement of A.muciniphilla treat group in intestinal immune response through hepatic RNA gene analysis. 32

6. Expression of serotonin receptor in the liver was improved by treatment of A.muciniphila. 34

7. Severity of patients with cirrhosis was correlated with the amount of portal vein serotonin. 36

Ⅳ. Disccusion 38

Ⅴ. References 42

Abstract Korean 45

Table 1. Primers used in the study. 22

Table 2. Antibodies used in the study. 22

Table 3. Flow diagram. Health Control, Cirrhosis groups divided by MELD Scroe 23

Table 4. Comparison of clinical parameters between healthy control, cirrhosis groups. 24

Supplemantion Data table1. Basic characteristics of human Portal blood subjects stratified by liver cirrhosis progression 45

Figure 1. Gut microbiota as altered by depending on severity in Cirrhosis patients. 25

Figure 2. Supplementation with A.muciniphilla ameliorates the progression of liver fibrosis 27

Figure 3. Supplementation with A.muciniphilla in humanized mouse model ameliorates the progression liver fibrosis better than conventional mouse model. 29

Figure 4. Mouse stool analysis in the DDC diet and A.muciniphilla supplementation model. 31

Figure 5. Results of QuantSeq mRNA-Seq analysis in DDC diet liver fibrosis model. 33

Figure 6. 5-HTR 2A/2B receptor expression are increased in DDC only treated group liver tissue, while in A.muciniphilla treated group 35

Figure 7. Human HVPG measurement patient biopsy and portal vein serum correlation with 5-HT. 37

Supplemantion Data1. Serotonin receptor qPCR data and Cardiac blood serotonin concentration 47