Role of intrinsic coagulation pathway in the pathogenesis of idiopathic pulmonary fibrosis [Elektronische Ressource] / vorgelegt von Jabłońska, Ewa Danuta

Jablonska, Ewa Danuta

Sujets

• Biologie

Informations

• Publié par: justus-liebig-universitat_giessen
• Publié le: 01 janvier 2011
• Nombre de lectures: 34
• Langue: English
• Poids de l'ouvrage: 6 Mo

Extrait

EWA JABLONSKA
Role of Intrinsic Coagulation Pathway in the
Pathogenesis of Idiopathic Pulmonary Fibrosis
édition scientifique
VVB LAUFERSWEILER VERLAG INAUGURALDISSERTATION zur Erlangung des Grades eines Doktors der Humanbiologie
VVB LAUFERSWEILER VERLAG des Fachbereichs Medizin der Justus-Liebig-Universität Gießen
STAUFENBERGRING 15 ISBN: 978-3-8359-5693-3
D-35396 GIESSEN
Tel: [phone] Fax: -5599890
[e-mail]
www.doktorverlag.de 9 7 8 3 8 3 5 9 5 6 9 3 3
édition scientifique
VVB LAUFERSWEILER VERLAGVVB
© Sebastian Kaulitzki - Fotolia.com
EWA JABLONSKA ROLE OF FXII IN THE PATHOGENESIS OF IPF
. Das Werk ist in allen seinen Teilen urheberrechtlich geschützt.
Jede Verwertung ist ohne schriftliche Zustimmung des Autors
oder des Verlages unzulässig. Das gilt insbesondere für
Vervielfältigungen, Übersetzungen, Mikroverfilmungen
und die Einspeicherung in und Verarbeitung durch
elektronische Systeme.
1. Auflage 2010
All rights reserved. No part of this publication may be
reproduced, stored in a retrieval system, or transmitted,
in any form or by any means, electronic, mechanical,
photocopying, recording, or otherwise, without the prior
written permission of the Author or the Publishers.
st1 Edition 2010
© 2010 by VVB LAUFERSWEILER VERLAG, Giessen
Printed in Germany
édition scientifique
VVB LAUFERSWEILER VERLAG
STAUFENBERGRING 15, D-35396 GIESSEN
Tel: [phone] Fax: [phone]
email: [e-mail]
www.doktorverlag.de


Role of Intrinsic Coagulation Pathway in the Pathogenesis of Idiopathic
Pulmonary Fibrosis






Inauguraldissertation
zur Erlangung des Grades eines Doktors der Humanbiologie
des Fachbereichs Medizin
der Justus-Liebig-Universität Gießen




vorgelegt von
Jab łońska, Ewa Danuta
aus
Toru ń, Polen


Giessen 2010 Aus dem Institut für Biochemie
des Fachbereichs Medizin der Justus-Liebig-Universität Giessen
Leiter/Direktor: Prof. Dr. Klaus T. Preissner























Gutachter: Prof. Dr. K. T. Preissner rof. Dr. W. Kummer

Tag der Disputation: 29.11.10.Table of contents
________________________________________________________________________
I. Table of contents
I. Table of contents...............................................................................................................I
II. List of figures ............................................................................................................... IV
III. List of tables.... VI
IV. List of abbreviations ..................................................................................................VII
V. Summary ....................................................................................................................... X
VI. Zusammenfassung......................................................................................................XII
1. Introduction..................................................................................................................... 1
1.1. Blood Coagulation Pathways ................................................................................... 1
1.1.1. Structural and functional characteristics of the intrinsic coagulation pathway
factors.......................................................................................................................... 1
1.1.1.1. Factor XII ................................................................................................... 1
1.1.1.2. Factor XI .................................................................................................... 3
1.1.1.3. High molecular weight kininogen.............................................................. 4
1.1.1.4. Kallikrein 5
1.1.2. Activation of FXII............................................................................................. 6
1.1.2.1. Contact activation of FXII ......................................................................... 6
1.1.2.2. Activation of the contact system on the endothelial cell surface ............... 7
1.1.2.3. Inhibition of FXII activity.......................................................................... 7
1.1.2.3.1. Inhibition of FXII activity in vitro ...................................................... 7
1.1.2.3.2. Inhibition of FXII activity in vivo ....................................................... 8
1.1.3. Physiologic activities of FXII ........................................................................... 9
1.1.3.1. FXII and inflammatory reactions ............................................................... 9
1.1.3.1.1. FXII in hereditary angioedema ........................................................... 9
1.1.3.1.2. FXII in sepsis ...................................................................................... 9
1.1.3.2. Factor XII in thrombosis .......................................................................... 10
1.1.3.3. Factor XII in fibrinolysis 11
1.1.3.4. Mitogenic activities of FXII..................................................................... 11
1.1.4. Characterization of FXII promoter.................................................................. 12
1.2. Idiopathic pulmonary fibrosis ................................................................................ 12
1.2.1. Role of Transforming Growth Factor-  in the pathogenesis of lung fibrosis..13
ITable of contents
________________________________________________________________________
1.2.2. Bleomycin model of lung fibrosis................................................................... 14
1.2.3. Role of coagulation in the pathogenesis of idiopathic pulmonary fibrosis..... 15
2. Aim of the study............................................................................................................ 17
3. Materials and methods .................................................................................................. 18
3.1. Materials................................................................................................................. 18
3.1.1. Equipment ....................................................................................................... 18
3.1.2. Reagents .......................................................................................................... 19
3.2. Methods.................................................................................................................. 21
3.2.1. Intratracheal bleomycin administration........................................................... 21
3.2.2. Pulmonary compliance measurements............................................................ 21
3.2.3. Lung preparation ............................................................................................. 22
3.2.4. Isolation of murine and human lung fibroblasts and cell culture.................... 22
3.2.5. Microdissection of lung tissue and alveolar epithelial type II cells ................ 23
3.2.6. RNA isolation and reverse transcriptase reaction ........................................... 23
3.2.7. Real Time PCR ............................................................................................... 24
3.2.8. Protein isolation and quantification ................................................................ 25
3.2.9. SDS polyacrylamide gel electrophoresis ........................................................ 25
3.2.10. Immunoblotting............................................................................................. 25
3.2.11. Immunocytochemistry................................................................................... 26
3.2.12. Immunohistochemistry.................................................................................. 27
3.2.13. Proliferation assay......................................................................................... 27
3.2.14. Immunoprecipitation..................................................................................... 28
3.2.15. Generation of FXII promoter constructs and site-directed mutagenesis....... 28
3.2.16. Transient transfection and luciferase assay................................................... 29
3.2.17. Antisense Oligonucleotides........................................................................... 29
3.2.18. Chromatin immunoprecipitation ................................................................... 29
3.2.19. Streptavidin pull-down assay ........................................................................ 30
3.2.20. Statistics ........................................................................................................ 31
4. Results........................................................................................................................... 32
4.1. Expression of FXII, FXI and HMWK is altered in idiopathic pulmonary fibrosis
lungs .............................................................................................................................. 32 <