[Role of Transforming Growth Factor Beta And Microtubule Instability In Fibrotic Disease]

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Acknowledgement

I would take this opportunity to thank my research supervisor, family and friends for their support and guidance without which this research would not have been possible.

DECLARATION

I, [type your full first names and surname here], declare that the contents of this dissertation/thesis represent my own unaided work, and that the dissertation/thesis has not previously been submitted for academic examination towards any qualification. Furthermore, it represents my own opinions and not necessarily those of the University.

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Abstract

The TGF-ß (transforming growth factor-ß) system signals via protein kinase receptors and Smad mediators to regulate a plethora of biological processes, including morphogenesis, embryonic development, adult stem cell differentiation, immune regulation, wound healing and inflammation. In addition, alterations of specific components of the TGF-ß signalling pathway may contribute to a broad range of pathologies such as cancer, cardiovascular pathology, fibrosis and congenital diseases. The knowledge about the mechanisms involved in TGF-ß signal transduction has allowed a better understanding of the disease pathogenicity as well as the identification of several molecular targets with great potential in therapeutic interventions.

Table of Content

CHAPTER I : INTRODUCTION1

Fibrotic Disease1

Dependent and Independent TGF-Beta Smad Pathways2

c-Able Pathway4

Rho-ROCK Pathway4

Microtubule-Associated Proteins6

Kinesin10

Dynein10

CHAPTER II: LITERATURE REVIEW11

CV system11

Atherosclerosis13

MGP Matrix13

Activation of the RAAS14

CHAPTER III : METHODOLOGY16

Cell Culture And Transfection16

Preparation Of Samples For Imaging17

Total internal Reflection Microscopy (TIRF)17

Time Lapsing Experiment17

Fiji IMAGING Software18

Using the Trajectory Macro 0.5 software18

CHAPTER IV: RESULTS AND DISCUSSION20

CHAPTER V: CONCLUSION38

REFERENCES42

CHAPTER I : INTRODUCTION

Fibrotic Disease

Fibrosis is a pathologic process, which includes scar formation and over production of extracellular matrix, by the connective tissue, as a response to tissue damage. The molecular process is not different from normal formation of connective tissue and extracellular matrix in the normal organs. The context, the environment and the over production make the difference. Fibrosis formation includes interaction between many cell types and cytokines, and when the balance becomes profibrotic, there is fibrosis formation.

Major profibrotic agents are type 2 CD4 positive lymphocytes, CD40 receptor and ligand interaction, and the following cytokines: IL-4, transforming growth factor b, platelet derived growth factor. The major antifibrotic agent is interferon gamma. Pathologies include: in the skin pathologic scarring as colloid and hypertrophic scar, cirrhosis of liver and gallbladder, in the heart and the kidneys, pulmonary and bone-marrow fibrosis, and scleroderma. Scleroderma is chronic connective tissue disease, expressed clinically by systemic sclerosis and diffuses fibrosis of the skin and viscera.

This is a progressive degenerative disorder of the blood vessels, skin, lungs, kidneys, heart and GI tract and for this reason this disease plays a major role in fibrosis research. Fibrosis is considered an irreversible process, at least clinically, and is usually treated by anti-inflammatory and immunosuppressive agents. This kind of therapy was not proven successful and sometimes it harms more than cures. Many patients suffer from fibrotic diseases and the aim is to develop anti-fibrotic agents, targeted to the pathologic molecular process. Progressing step by step in this issue has direct clinic affect.

Dependent and Independent TGF-Beta Smad Pathways

A requirement for Smad signaling in ...