282 317

Cited 0 times in

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

Other Title
섬유성 망막 질환에서 전이성장인자 신호전달과정 억제를 통한 항섬유화 약물 (pirfenidone)의 효과 및 기전
Authors
이, 기황
Department
대학원 의학과
Degree
Doctor (2015)
Table Of Contents
I. 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
Abstract
Purpose: 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.
Keywords
Transforming growth factor-β (TGF-β)retinal pigment epithelial (RPE) cellsfibrosispirfenidonecell signalepithelial-mesenchymal transition전이성장인자망막색소상피섬유화퍼페니돈세포신호상피세포의 간질세포로의 변이
Appears in Collections:
Theses > School of Medicine / Graduate School of Medicine > Doctor
AJOU Authors
이, 기황
Full Text Link
Files in This Item:
000000020173.pdfDownload
Export
RIS (EndNote)
XLS (Excel)
XML

qrcode

해당 아이템을 이메일로 공유하기 원하시면 인증을 거치시기 바랍니다.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse