표제지
목차
국문 요약문 12
약어목록 15
Ⅰ. 서론 16
Ⅱ. 문헌 고찰 20
1. 염증성 장질환(Inflammatory bowel disease, IBD)의 발생 현황과 관리 20
2. 염증성 장질환과 장내 미생물의 관련성 23
3. 염증성 장질환에서 prebiotics의 역할 25
(1) Prebiotics 개론 25
(2) N-acetylglucosamine(NAG)의 적용 29
Ⅲ. 연구 내용 및 방법 31
1. 연구 디자인 31
2. 시험 시료 및 식이 제조 33
3. DAI(Disease Activity Index) 측정 33
4. 대장 조직병리학적 분석 33
1) Hematoxylin & eosin 염색 34
2) Alcian blue 염색 34
5. Intestinal permeability 평가 34
1) 혈청 FITC 농도 측정 34
2) 혈청 CD14 농도 측정 35
6. 대장 조직 단백질 추출 및 발현 35
7. 대장 조직 mRNA 추출 및 발현 37
8. 장내 미생물 조성 분석 37
9. 통계 분석 38
Ⅳ. 결과 39
1. 체중 변화 39
2. 식이 섭취량 41
3. 대장 길이 비교 43
4. Disease Activity Index 45
5. 대장 조직병리학적 분석 47
1) 조직 손상도 평가(Rectal histological score) 47
2) Mucus layer integrity 확인 52
3) Immunohistochemical quantification of CDX2 56
4) Immunohistochemical quantification of MUC2 59
6. Intestinal permeability 확인 62
1) Serum FITC concentration 62
2) Serum CD14 concentration 64
7. 대장 조직 단백질 발현 분석 66
1) Tight junction proteins 66
2) Inflammation-asssociated markers 69
8. 대장 조직 mRNA 발현 분석 85
1) 대장 조직 TNF-α 85
9. 분변 마이크로비오타 분석 88
1) 장내 미생물 다양성 88
2) PCoA 91
3) 장내 미생물 구성 93
4) Linear Discriminant Analysis(LDA) 108
5) 장내 미생물과 biomarker 상관관계 분석 112
6) KEGG metabolic pathway 분석 114
Ⅴ. 고찰 117
Ⅵ. 요약 및 결론 128
Ⅶ. 참고문헌 131
ABSTRACT 146
Table 1. Final body weight 40
Table 2. Average food intake 42
Table 3. Colon length of the experimental animals 44
Table 4. Final DAI score 46
Table 5. Histological score of the rectum 51
Table 6. Alcian blue intensity and goblet cell counts 55
Table 7. CDX2 staining in mouse intestine tissue 58
Table 8. MUC2 staining in mouse intestine tissue 61
Table 9. Serum concentration of FITC 63
Table 10. Serum concentration of CD14 65
Table 11. Colonic occludin, claudin-1 and ZO-1 protein expressions 68
Table 12. Colonic T-bet, GATA3, Foxp3 and RORγt protein expressions 71
Table 13. Colonic IFN-γ proteins expressions 73
Table 14. Colonic smad proteins expressions 76
Table 15. Colonic TGF-β1 protein expressions 78
Table 16. Colonic COX-2 protein expressions 81
Table 17. Colonic P-IκBα protein expressions 84
Table 18. Colonic TNF-α mRNA expressions 87
Table 19. Alpha-diversity of gut microbiota 90
Table 20. Firmicutes/Bacteroidetes ratio 97
Table 21. Correlation between gut microbiota and biomarkers 113
Figure 1. Hypothesis of the study 19
Figure 2. Increase in worldwide incidence of ulcerative colitis over time 22
Figure 3. Accepted Prebiotics and Candiates 27
Figure 4. Key functions of Prebiotics 28
Figure 5. Experimental design of study 32
Figure 6. Body weight changes during the experimental period 39
Figure 7. Average food intake during the experimental period 41
Figure 8. Colon length of the experimental animals 43
Figure 9. DAI score changes during the experimental period 45
Figure 10. H&E staining of the rectum and pathological scoring criteria 48
Figure 11. Representative histological features of the rectum 49
Figure 12. Histological score of the rectum 50
Figure 13. Representative Alcian blue staining features of rectum 53
Figure 14. Alcian blue intensity of rectum 54
Figure 15. Representative images of CDX2 immunohistochemistry of colon tissue 57
Figure 16. Representative images of MUC2 immunohistochemistry of colon tissue (magnification × 50) 60
Figure 17. Serum FITC concentration 62
Figure 18. Serum CD14 concentration 64
Figure 19. Colonic occludin, claudin-1 and ZO-1 protein expressions 67
Figure 20. Colonic T-bet, GATA3, Foxp3, RORγt protein expressions 70
Figure 21. Colonic IFN-γ proteins expressions 72
Figure 22. Colonic smad proteins expressions 75
Figure 23. Colonic TGF-β1 protein expressions 77
Figure 24. Colonic COX-2 protein expressions 80
Figure 25. Colonic P-IκBα protein expressions 83
Figure 26. Colonic TNF-α mRNA expressions 86
Figure 27. Alpha-diversity of gut microbiota 89
Figure 28. Beta-diversity of gut microbiota 92
Figure 29. Relative abundance of gut microbiota at phylum level 94
Figure 30. Composition of gut microbiota at phylum level 95
Figure 31. Firmicutes/Bacteroidetes ratio 96
Figure 32. Relative abundance of gut microbiota at class level 99
Figure 33. Relative abundance of gut microbiota at order level 101
Figure 34. Relative abundance of gut microbiota at family level 103
Figure 35. Relative abundance of gut microbiota at genus level 105
Figure 36. Relative abundance of gut microbiota at species level 107
Figure 37. LDA score of each exper imental group(continued) 110
Figure 38. Heatmap of predicted functional KEGG pathways(continued) 115