Molecular mechanisms for the inhibition of the nuclear factor {_k63B [kappa-B] signal transduction in intestinal epithelial cells under conditions of chronic inflammation [Elektronische Ressource] / Pedro Antonio Ruiz-Castro

Cacahuete

Sujets

• Gesundheit

Informations

• Publié par: technische_universitat_munchen
• Publié le: 01 janvier 2007
• Nombre de lectures: 16
• Langue: English
• Poids de l'ouvrage: 7 Mo

Extrait

Fachgebiet für Experimentelle Ernährungsmedizin



Molecular mechanisms for the inhibition of the nuclear factor
κB signal transduction in intestinal epithelial cells under
conditions of chronic inflammation



Pedro Antonio Ruiz-Castro



Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum Weihenstephan für
Ernährung, Landnutzung und Umwelt der Technischen Universität München zur
Erlangung des akademischen Grades eines

Doktors der Naturwissenschaften

genehmigten Dissertation.



Vorsitzender: Univ.-Prof. Dr. Michael Schemann

Prüfer der Dissertation: 1. Univ.-Prof. Dr. Dirk Haller
2. Univ.-Prof. Dr. Hannelore Daniel
3. Univ.-Prof. Dr. Roland M. Schmid


Die Dissertation wurde am 08.02.2007 bei der Technischen Universität München
eingereicht und durch die Fakultät für Ernährung, Landnutzung und Umwelt am
30.07.2007 angenommen.









































A mis padres,
Rosa María Castro Abad y Pedro Ruiz Gil
por su constante ánimo y apoyo


The presented thesis is based on the following peer-reviewed original papers and
submitted manuscripts:

I. Ruiz, PA, Kim, SC, Sartor, RB and Haller, D
“15-deoxy-delta12,14-prostaglandin J2-mediated ERK signaling inhibits gram-
negative bacteria-induced RelA phosphorylation and interleukin-6 gene
expression in intestinal epithelial cells through modulation of protein phosphatase
2A activity” J. Biol. Chem., 2004, Aug 20; 279(34):36103-11

II. Ruiz, PA*, Shkoda, A*, Kim, SC, Sartor, RB and Haller, D
“IL-10 gene-deficient mice lack TGF- β/Smad signaling and fail to inhibit pro-
inflammatory gene expression in intestinal epithelial cells after the colonization
with colitogenic Enterococcus faecalis” J. Immunol., 2005, 174: 2990-2999
*Authors contributed equally

III. Ruiz, PA, Hoffmann, M, Szcesny, S, Blaut, M and Haller, D
“Innate mechanisms for Bifidobacterium lactis to activate transient pro-
inflammatory host responses in intestinal epithelial cells after the colonization of
germfree rats” Immunology, 2005, 115, 441-450

IV. Ruiz, PA and Haller, D
"Functional Diversity of Flavonoids in the Inhibition of the Pro-inflammatory NF-
κB, IRF, and Akt Signaling Pathways in Murine Intestinal Epithelial Cells" J.
Nutr., 2006, 136:664-671

V. Shkoda, A*, Ruiz, PA*, Daniel, H, Kim, SC, Rogler, G, Sartor, RB and Haller, D
“Interleukin 10 blocked endoplasmic reticulum stress in intestinal epithelial cells:
impact on chronic inflammation” Gastroenterology, 2007, 132(1): 190-207
*Authors contributed equally

VI. Ruiz, PA, Braune, A, Hölzlwimmer, G, Quintanilla-Fend and Haller, D
“Quercetin inhibits TNF-induced NF-κB transcription factor recruitment to pro-
inflammatory gene promoters in murine intestinal epithelial cells” Submitted to J.
Nutr.


















“What are the facts? Again and again and again—what are the facts? Shun
wishful thinking, ignore divine revelation, forget what "the stars foretell," avoid

opinion, care not what the neighbors think, never mind the unguessable
"verdict of history"—what are the facts, and to how many decimal places? You
pilot always into an unknown future; facts are your single clue. Get the facts!”

Robert A. Heinlein
(1907-1988)
Table of Contents vi

TABLE OF CONTENTS
SUMMARY ....................................................................................................................... 9
PROLOGUE ................................................................................................................... 10
INTRODUCTION ............................................................................................................ 11
1 Ecology of the gastrointestinal tract .......................................................................... 11
1.1 Host-microbial interactions.............................................................................. 11
1.2 Assembling of the intestinal mucosa............................................................... 13
1.3 Gut-associated lymphoid tissue ...................................................................... 14
2 Inflammatory bowel disease (IBD) ............................................................................ 16
2.1 Models of mucosal inflammation ..................................................................... 17
2.2 Probiotics and IBD........................................................................................... 19
2.3 Terapeutic role of flavonoids in IBD ................................................................ 22
3 IECs and the mucosal immune system 24
4 NF-κB signaling......................................................................................................... 26
4.1 Rel/NF-κB and IκBα families........................................................................... 28
4.2 NF-κB signaling pathway ................................................................................ 30
5 TLR-mediated NF-κB activation 34
5.1 TLR4................................................................................................................ 36
5.2 TLR2 36
5.3 TLR-mediated signaling pathways .................................................................. 37
6 MAPKs ...................................................................................................................... 39
7 Cytokine signaling and IBD....................................................................................... 41
7.1 TGF-β/Smad signaling .................................................................................... 41
7.2 IL-10 signaling pathway................................................................................... 43
AIMS OF THE WORK .................................................................................................... 46
RESULTS AND DISCUSSION 48
12,141 15-deoxy-Δ -prostaglandin J inhibits Gram-negative bacteria-induced 2
pro-inflammatory responses in IECs through modulation of protein
phosphatase 2A activity ............................................................................................ 48
Table of Contents vii

2 Mechanisms for Bifidobacterium lactis BB12-induced transient pro-
inflammatory processes in the intestinal epithelium.................................................. 54
3 TGF-β1 inhibits TLR2-mediated Enterococcus faecalis induction of NF-κB
signaling pathway in IECs......................................................................................... 58
4 IL-10 abrogates ER stress response by blocking ATF-6 recruitment to the
Grp78 promoter......................................................................................................... 61
5 Functional diversity of flavonoids and its bacterial metabolites in the
inhibition of pro-inflammatory signaling pathways in IECs ........................................ 66
ZUSAMMENFASSUNG.................................................................................................. 74
REFERENCES ............................................................................................................... 75
LIST OF ABBREVIATIONS.......................................................................................... 103
LIST OF FIGURES ....................................................................................................... 107
LIST OF TABLES......................................................................................................... 108
APPENDIX 1................................................................................................................. 110
APPENDIX 2 120
APPENDIX 3 131
APPENDIX 4 142
APPENDIX 5 151
APPENDIX 6 170
ACKNOWLEDGEMENTS............................................................................................. 180
CURRICULUM VITAE .................................................................................................. 182






























Summary 9

SUMMARY
Intestinal bowel disease (IBD) is an inflammatory disorder which results from an
exacerbated mucosal immune response induced by the indigenous microbiota. The
nuclear factor (NF)-κB is responsible for the expression of many genes involved in
immunity and inflammation. In the present work, animal models as well as cell lines were
used to characterize the molecular mechanisms for the downregulation of the NF-κB
signalling pathway in intestinal epithelial cells (IECs), which play a crucial role in mucosal