Title Page
ABSTRACT
Contents
LIST OF ABBREVIATIONS 11
INTRODUCTION 12
MATERIALS AND METHODS 15
Study design 15
Peritoneal adhesion induction 17
Peritoneal adhesion evaluation 19
Histologic examination 20
Quantitative real-time polymerase chain reaction (qRT-PCR) analysis 21
Western blot analysis 22
Immunohistochemical analysis 23
Immunofluorescence analysis 24
Statistical analysis 25
RESULTS 26
EW-7197 decreased the incidence and severity of peritoneal adhesions 26
Fibrosis and collagen deposition were attenuated in EW-7197 treated peritoneal ischemic buttons 30
TGF-β/Smad2/3 signaling and expression of CTGF and VEGF were reduced in EW-7197 treated peritoneal ischemic buttons 36
Markers of MMT and ECM synthesis and degradation were altered in EW-7197 treated peritoneal ischemic buttons 40
MMT was inhibited in a Smad-dependent manner in EW-7197 treated peritoneal ischemic buttons 46
DISCUSSION 47
REFERENCES 51
Table 1. Adhesion Score 19
Table 2. Fibrosis and inflammation grading scale 20
Table 3. List of primers Used for qRT-PCR 21
Fig. 1. A flow diagram showing randomization process and study follow-up. 16
Fig. 2. Photograph shows four ischemic buttons (arrows) created with 4-0 silk suture (Ailee,... 18
Fig. 3. Bar graph of incidence of peritoneal adhesions. 27
Fig. 4. Bar graph of quality score of peritoneal adhesions. 27
Fig. 5. Bar graph of tenacity score of peritoneal adhesions. 28
Fig. 6. Photograph shows severe peritoneal adhesion formation (arrows = 3 quality score;... 28
Fig. 7. Photograph shows moderate peritoneal adhesion formation (arrows = 1 quality score;... 29
Fig. 8. Photograph shows mild peritoneal adhesion formation (arrows = 1 quality score) in... 29
Fig. 9. Bar graph of fibrosis score of peritoneal ischemic buttons. 31
Fig. 10. Bar graph of inflammation score of peritoneal ischemic buttons. 31
Fig. 11. Bar graph of collagen deposition score of peritoneal ischemic buttons. 32
Fig. 12. Histologic section of a peritoneal ischemic button from control group shows dense... 32
Fig. 13. Histologic section of a peritoneal ischemic button from 10 mg EW-7197 group... 33
Fig. 14. Histologic section of a peritoneal ischemic button from 20 mg EW-7197 group... 33
Fig. 15. Histologic section of a peritoneal ischemic button from control group shows... 34
Fig. 16. Histologic section of a peritoneal ischemic button from 10 mg EW-7197 group... 34
Fig. 17. Histologic section of a peritoneal ischemic button from 20 mg EW-7197 group... 35
Fig. 18. Bar graph of mean mRNA expression of TGF-β 37
Fig. 19. Bar graph of mean protein expression of Smad2/3 with representative bands 37
Fig. 20. Bar graph of mean protein expression of p-Smad2/3 with representative bands 38
Fig. 21. Bar graph of mean mRNA expression of CTGF 38
Fig. 22. Bar graph of mean mRNA expression of VEGF 39
Fig. 23. Bar graph of mean mRNA expression of Snail-1 41
Fig. 24. Bar graph of mean protein expression of Snail-1 with representative bands 42
Fig. 25. Bar graph of mean mRNA expression of E-Cadherin 42
Fig. 26. Bar graph of mean mRNA expression of α-SMA 43
Fig. 27. Bar graph of mean mRNA expression of COL-1 43
Fig. 28. Bar graph of mean protein expression of COL-1 with representative bands 44
Fig. 29. Bar graph of mean mRNA expression of TIMP-1 44
Fig. 30. Bar graph of mean mRNA expression of TIMP-1 45
Fig. 31. Immunofluorescence (DAPI = blue; E-Cadherin = green; α-SMA = red) and... 46