Haaften, . W.T. van (2012) In vitro model to unravel the mechanism of intestinal fibrosis in IBD. thesis, Medicine.
Full text available on request.Abstract
Background and aims: Intestinal fibrosis is a clinically important complication in inflammatory bowel disease (IBD) resulting from excessive deposition of extracellular matrix in reaction to chronic inflammation. Important contributors to this process could be stellate cells and heat shock protein 47 (HSP47). In liver, the stellate cell is known to play a major role in fibrosis. Synaptophysin is said to be a marker for stellate cells. HSP47 is involved maturation and secretion of collagen. The aim of this study is to investigate the role of stellate cells and HSP47 in vitro using precision-cut intestinal slices (PCIS). Methods: Intestinal slices were obtained from rat, and human (healthy and suffering from Crohn’s disease (CD)) intestine and incubated up to 72h with TGF-β (5ng/ml). Afterwards, ATP content, Park score, and gene expression of fibrosis markers was determined. Immunohistochemical staining for HSP47 and synaptophysin in healthy and CD intestinal tissue, and for collagen-I in incubated slices were also performed. Results: Rat and human intestinal slices remained viable during incubation. Incubation with TGF-β did not show any significant differences in viability compared to control. Gene expression of HSP47 and synaptophysin increased during incubation of human PCIS. In contrast to Alpha-smooth muscle actin, which decreased and Pro-col-1a1 that remained constant during culture. That is in agreement with the findings from the immunohistochemical staining of incubated slices for Collagen-I; protein of Collagen-1 also remained constant. Immunohistochemical staining showed that the expression of synaptophysin was higher in healthy intestine and the expression of HSP47 did not differ compared to intestine of CD patients. Conclusions: A reproducible technique to induce the onset of the process of fibrosis was developed. Based on these findings, the presence of the intestinal stellate cell and its involvement in the process of fibrosis and the production of HSP47 are plausible. Double staining for synaptophysin and HSP47, and for synaptophysin and other stellate cell markers should be performed to confirm this
| Item Type: | Thesis (Thesis) |
|---|---|
| Supervisor name: | Faculty supervisor: and Leuvenink, Dr. H.G.D. PhD |
| Supervisor name: | External supervisor: and Olinga, Dr. P.PhD and Daily supervisor: and Pham, MSc B.T. Pharmaceutical Technology and Biopharmacy |
| Faculty: | Medical Sciences |
| Date Deposited: | 25 Jun 2020 10:51 |
| Last Modified: | 25 Jun 2020 10:51 |
| URI: | https://umcg.studenttheses.ub.rug.nl/id/eprint/1238 |
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