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Effect of Anti-fibrotic Drug (Pirfenidone) on TGF-β Signaling Pathway in Ocular Fibrotic Disorders

DC Field Value Language
dc.contributor.author이, 기황-
dc.date.accessioned2015-11-03T04:09:13Z-
dc.date.available2015-11-03T04:09:13Z-
dc.date.issued2015-
dc.identifier.urihttp://repository.ajou.ac.kr/handle/201003/11886-
dc.description.abstractPurpose: Transforming growth factor-β (TGF-β) plays a key role in transforming retinal pigment epithelial (RPE) cells into mesenchymal fibroblastic cells, which are implicated in fibrotic disorders of the retina. Herein, the effect of pirfenidone, a novel anti-fibrotic agent, on TGF-β1-mediated fibrogenesis in the human RPE cell line (ARPE)-19 was tested.

Methods: The effect of pirfenidone on the TGF-β1-induced phenotype in ARPE-19 cells was measured with immunocytochemistry as the change in F-actin. Fibronectin and collagen production was measured with enzyme-linked immunosorbent assay, and cell migration activity was investigated using a scratch assay. Immunoblot analyses of cofilin, Smad protein (Smad) 2/3, p38 mitogen-activated protein kinase, c-Jun N-terminal kinase, and extracellular signal related kinase expression were conducted to elucidate the cell signaling networks that contribute to the anti-fibrotic effect of pirfenidone.

Results: Treatment with TGF-β1 induced typical phenotypic changes such as formation of stress fiber running parallel to the long axis of cells, and enhanced migration and production of extracellular matrix components such as collagen type I and fibronectin. This fibroblast-like phenotype induced by TGF-β1 was significantly inhibited by pretreatment with pirfenidone in a dose-dependent manner. Pirfenidone inhibited TGF-β signaling by preventing nuclear accumulation of active Smad2/3 complexes rather than phosphorylation of Smad2/3.

Conclusions: These results collectively provide a rational background for future evaluation of pirfenidone as a potential anti-fibrotic agent for treating proliferative vitreoretinopathy and other fibrotic retinal disorders.
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dc.description.tableofcontentsI. INTRODUCTION 1

A. Fibrotic Retinal Diseases 1

B. Retinal Pigment Epithelial Cells 2

C. Epithelia-to- Mesenchymal Transition 3

D. Transforming Growth Factor 4

1. Smad-Dependent Signaling 4

2. Smad-Independent Signaling 5

E. Novel Anti-fibrotic Drug (Pirfenidone) 5

F. Aims of This Study 8

II. MATERIALS AND METHODS 9

A. MATERIALS 9

1.Reagents 9

B. METHODS 9

1. Cell Cuture 9

2. Enzyme-linked Immunosorbent Assay 9

3. Immunocytochemistry 10

4. Cell Migration Assay 10

5. Immunoblot analysis 10

6. Statistical analysis 11

III. RESULTS 12

A. Pirfenidone inhibits TGF-β1 induced fibroblastic phenotypes in ARPE-19 cells 12

B. Pirfenidone suppresses the TGF-β1-induced expression of ECM components in ARPE-19 cells 22

C. Pirfenidone abrogates the TFG-β1-induced migration of ARPE-19 cells 22

D. Pirfenidone blocks TGF-β1-induced nuclear translocation but not phosphorylation of Smads 27

IV. DISCUSSION 33

A. Pirfenidone and TGF-β1 induced fibroblastic phenotypes in ARPE-19 cells 33

B. Pirfenidone and TGF-β1-induced expression of extracellular matrix components in ARPE-19 cells 35

C. Possible molecular mechanisms of pirfenidone for the inhibitory action for TGF-β1-induced fibrogenesis in ARPE-19 cells 36

V. CONCLUSION 41

REFERENCES 42

국문요약 50
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dc.language.isoen-
dc.titleEffect of Anti-fibrotic Drug (Pirfenidone) on TGF-β Signaling Pathway in Ocular Fibrotic Disorders-
dc.title.alternative섬유성 망막 질환에서 전이성장인자 신호전달과정 억제를 통한 항섬유화 약물 (pirfenidone)의 효과 및 기전-
dc.typeThesis-
dc.identifier.urlhttp://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000020173-
dc.subject.keywordTransforming growth factor-β (TGF-β)-
dc.subject.keywordretinal pigment epithelial (RPE) cells-
dc.subject.keywordfibrosis-
dc.subject.keywordpirfenidone-
dc.subject.keywordcell signal-
dc.subject.keywordepithelial-mesenchymal transition-
dc.subject.keyword전이성장인자-
dc.subject.keyword망막색소상피-
dc.subject.keyword섬유화-
dc.subject.keyword퍼페니돈-
dc.subject.keyword세포신호-
dc.subject.keyword상피세포의 간질세포로의 변이-
dc.description.degreeDoctor-
dc.contributor.department대학원 의학과-
dc.contributor.affiliatedAuthor이, 기황-
dc.date.awarded2015-
dc.type.localTheses-
dc.citation.date2015-
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