Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis:
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| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
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Münster
2022
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| Online-Zugang: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Beschreibung: | XII, 122 Blätter Illustrationen, Diagramme 30 cm |
Internformat
MARC
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| 100 | 1 | |a Schulz, Annika |d 1992- |e Verfasser |0 (DE-588)1275034012 |4 aut | |
| 245 | 1 | 0 | |a Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis |c vorgelegt von Annika Schulz aus Thuine |
| 264 | 1 | |a Münster |c 2022 | |
| 300 | |a XII, 122 Blätter |b Illustrationen, Diagramme |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Westfälische Wilhelms-Universität Münster |d 2022 |g Beschränkt für den Austausch | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | 2 | |m B:DE-101 |q application/pdf |u https://d-nb.info/1275742602/04 |3 Inhaltsverzeichnis |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034225268&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-034225268 | ||
| 883 | 0 | |8 1\p |a emakn |c 0,04785 |d 20230329 |q DE-101 |u https://d-nb.info/provenance/plan#emakn | |
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| 883 | 2 | |8 3\p |a dnb |d 20230330 |q DE-101 |u https://d-nb.info/provenance/plan#dnb | |
Datensatz im Suchindex
| _version_ | 1804185193874980864 |
|---|---|
| adam_text | TABLE
OF
CONTENTS
ABBREVIATION
REGISTER
...........................................................................................................
V
LIST
OF
FIGURES
........................................................................................................................
X
LIST
OF
TABLES
.......................................................................................................................
XII
ABSTRACTS
.................................................................................................................................
1
ABSTRAKT
(DEUTSCH)
............................................................................................................
1
ABSTRACT
(ENGLISH)
..............................................................................................................
3
1
INTRODUCTION
.........................................................................................................................
5
1.1
.
SOLUBLE
FMS-LIKE
TYROSINE
KINASE
.......................................................................
5
1.1.1
.
SFLT-1
AS
A
DECOY
RECEPTOR
FOR
VASCULAR
ENDOTHELIAL
GROWTH
FACTOR
.....
7
1.1.2
BINDING
TO
A5PI
INTEGRIN
FAVORS
ANGIOGENESIS
..................................................
8
1.2
THE
ENDOTHELIUM:
HOMEOSTASIS
VS.
DISEASE
............................................................
9
1.2.1
THE
ENDOTHELIAL
GLYCOCALYX
...............................................................................
11
1.2.2
ENDOTHELIAL
CORTICAL
STIFFNESS
.............................................................................
13
1.3
PATHWAYS
AND
MECHANISMS
INVOLVED
IN
ENDOTHELIAL
DYSFUNCTION
-
BRIEF
REMARKS
...........................................................................................................................................
15
1.3.1
OXIDATIVE
STRESS
.................................................................................................
15
1.3.2
P38
MITOGEN-ACTIVATED
PROTEIN
KINASE
PATHWAY
..............................................
16
1.3.3
ENDOTHELIAL-TO-MESENCHYMAL
TRANSITION
...........................................................
16
1.4
AIMS
OF
THE
STUDY
.......................................................................................................
17
2
MATERIAL
..............................................................................................................................
19
2.1
DEVICES
......................................................................................................................
19
2.2
CHEMICALS
AND
BIOLOGICAL
REAGENTS
.........................................................................
20
2.3
CONSUMABLES
............................................................................................................
22
2.4
BUFFERS,
SOLUTIONS,
AND
MEDIA
..................................................................................
24
2.5
SURGICAL
MATERIAL
.......................................................................................................
25
2.6
KITS
..............................................................................................................................
27
2.7
ANTIBODIES
..................................................................................................................
27
2.8
PRIMERS
.......................................................................................................................
28
2.9
SOFTWARE
.....................................................................................................................
29
3
METHODS
.............................................................................................................................
31
3.1
EXPERIMENTAL
DESIGN
.................................................................................................
31
3.2
CELL
CULTURE
AND
CELL
CULTURE-BASED
ASSAYS
............................................................
31
3.2.1
CULTIVATION
OF
PRIMARY
HUMAN
UMBILICAL
VEIN
ENDOTHELIAL
CELLS
.....................
31
3.2.2
CULTIVATION
OF
EA.HY926
....................................................................................
32
3.2.3
CULTIVATION
OF
THP-1
..........................................................................................
33
3.2.4
ATOMIC
FORCE
MICROSCOPY
..................................................................................34
3.2.5
DETERMINATION
OF
EGC
COMPONENTS
................................................................37
3.2.6
MONOCYTE-TO-ENDOTHELIAL-CELL
ADHESION
ASSAY
................................................38
3.2.7
TRANSMIGRATION
ASSAY
.......................................................................................39
3.2.8
ADHESION
MOLECULE
EXPRESSION
BY
FLOW
CYTOMETRY
........................................39
3.2.9
SDS-POLYACRYLAMIDE
GEL
ELECTROPHORESIS
.......................................................
40
3.2.10
WESTERN
BLOTTING
...............................................................................................
41
3.2.11
IN
SILICO
MOLECULAR
DOCKING
ANALYSIS
..............................................................
42
II
3.2.12
VIABILITY
ASSAY
..................................................................................................
43
3.2.13
F-ACTIN
BINDING
AND
STAINING
..........................................................................
43
3.2.14
ENDOTHELIAL
CELL
PERMEABILITY
..........................................................................
44
3.2.15
SCRATCH
ASSAY
..................................................................................................
45
3.2.16
CELLULAR
ROS
DETECTION
...................................................................................
45
3.2.17
RNA
ISOLATION
...................................................................................................
46
3.2.18
POLYMERASE
CHAIN
REACTION
............................................................................
47
3.3
ANIMAL
MODEL
............................................................................................................
51
3.3.1
SURGICAL
IMPLANTATION
OF
OSMOTIC
MINIPUMPS
..................................................52
3.3.2
ATOMIC
FORCE
MICROSCOPY
..................................................................................
53
3.3.3
TISSUE
FIBROSIS
DETERMINATION
...........................................................................
54
3.3.4
RNA
ISOLATION
......................................................................................................55
3.3.5
POLYMERASE
CHAIN
REACTION
...............................................................................
56
3.4
STATISTICAL
ANALYSIS
...................................................................................................
56
3
RESULTS
...............................................................................................................................
57
3.1
CONFORMATIONAL
CHANGES
OF
THE
ENDOTHELIAL
GLYCOCALYX
UNDER
EXCESS
SFLT-1
...
57
3.1.1
EXCESS
SFLT-1
LEADS
TO
CONFORMATIONAL
CHANGES
OF
THE
EGC
.......................
57
3.1.2
.
EXCESS
SFLT-1
INCREASES
MONOCYTE
ADHESION
.....................................
62
3.1.3
ENDOTHELIAL
CELL
VIABILITY
UNDER
SFLT-1
...............................................................
65
3.2
EXCESS
SFLT-1
AFFECTS
ENDOTHELIAL
STIFFNESS
.............................................................
69
3.2.1
EXCESS
SFLT-1
STIFFENS
ENDOTHELIAL
CELL
CORTEX
IN
VITRO
AND
IN
VIVO
................
69
3.2.2
SFLT-1
TREATMENT
ACTIVATES
P38
MAPK
STRESS
RESPONSE
PATHWAY
................
71
III
3.2.3
SFLT-1
INDUCES
CHANGES
IN
THE
F-ACTIN
DISTRIBUTION
AND
DISTURBS
THE
BARRIER
FUNCTION
OF
ENDOTHELIAL
CELLS
.......................................................................................
73
3.2.4
SFLT-1
ONLY
SLIGHTLY
AFFECTS
MECHANOSENSITIVE
GENES
IN
HUVECS
..............
76
3.2.5
SFLT-1
AFFECTS
MECHANOSENSITIVE
GENES
IN
THE
MICE
LUNG
..............................
78
5
DISCUSSION
........................................................................................................................
81
5.1
CONFORMATIONAL
CHANGES
OF
THE
ENDOTHELIAL
GLYCOCALYX
UNDER
EXCESS
SFLT-1
...
81
5.2
EXCESS
SFLT-1
AFFECTS
ENDOTHELIAL
STIFFNESS
.............................................................
87
5.3
LIMITATIONS
AND
REMARKS
...........................................................................................
92
5.4
CONCLUSION
AND
PERSPECTIVES
..................................................................................
93
6
APPENDIX
...........................................................................................................................
94
6.1
MONOCYTE
ADHESION
UNDER
EXCESS
SFLT-1
...............................................................
94
6.2
CELL
VIABILITY
UPON
SFLT-1
TREATMENT
.........................................................................95
6.3
CORTICAL
STIFFNESS
UNDER
TGF-32
TREATMENT
............................................................
96
6.4
P38
MAPK
INHIBITOR
SB203580
PREVENTS
COLLAPSE
OF
THE
EGC
............................
97
REFERENCES
..........................................................................................................................
99
CURRICULUM
VITAE
................................................................................................................119
DANKSAGUNG
......................................................................................................................122
IV
|
| adam_txt |
TABLE
OF
CONTENTS
ABBREVIATION
REGISTER
.
V
LIST
OF
FIGURES
.
X
LIST
OF
TABLES
.
XII
ABSTRACTS
.
1
ABSTRAKT
(DEUTSCH)
.
1
ABSTRACT
(ENGLISH)
.
3
1
INTRODUCTION
.
5
1.1
.
SOLUBLE
FMS-LIKE
TYROSINE
KINASE
.
5
1.1.1
.
SFLT-1
AS
A
DECOY
RECEPTOR
FOR
VASCULAR
ENDOTHELIAL
GROWTH
FACTOR
.
7
1.1.2
BINDING
TO
A5PI
INTEGRIN
FAVORS
ANGIOGENESIS
.
8
1.2
THE
ENDOTHELIUM:
HOMEOSTASIS
VS.
DISEASE
.
9
1.2.1
THE
ENDOTHELIAL
GLYCOCALYX
.
11
1.2.2
ENDOTHELIAL
CORTICAL
STIFFNESS
.
13
1.3
PATHWAYS
AND
MECHANISMS
INVOLVED
IN
ENDOTHELIAL
DYSFUNCTION
-
BRIEF
REMARKS
.
15
1.3.1
OXIDATIVE
STRESS
.
15
1.3.2
P38
MITOGEN-ACTIVATED
PROTEIN
KINASE
PATHWAY
.
16
1.3.3
ENDOTHELIAL-TO-MESENCHYMAL
TRANSITION
.
16
1.4
AIMS
OF
THE
STUDY
.
17
2
MATERIAL
.
19
2.1
DEVICES
.
19
2.2
CHEMICALS
AND
BIOLOGICAL
REAGENTS
.
20
2.3
CONSUMABLES
.
22
2.4
BUFFERS,
SOLUTIONS,
AND
MEDIA
.
24
2.5
SURGICAL
MATERIAL
.
25
2.6
KITS
.
27
2.7
ANTIBODIES
.
27
2.8
PRIMERS
.
28
2.9
SOFTWARE
.
29
3
METHODS
.
31
3.1
EXPERIMENTAL
DESIGN
.
31
3.2
CELL
CULTURE
AND
CELL
CULTURE-BASED
ASSAYS
.
31
3.2.1
CULTIVATION
OF
PRIMARY
HUMAN
UMBILICAL
VEIN
ENDOTHELIAL
CELLS
.
31
3.2.2
CULTIVATION
OF
EA.HY926
.
32
3.2.3
CULTIVATION
OF
THP-1
.
33
3.2.4
ATOMIC
FORCE
MICROSCOPY
.34
3.2.5
DETERMINATION
OF
EGC
COMPONENTS
.37
3.2.6
MONOCYTE-TO-ENDOTHELIAL-CELL
ADHESION
ASSAY
.38
3.2.7
TRANSMIGRATION
ASSAY
.39
3.2.8
ADHESION
MOLECULE
EXPRESSION
BY
FLOW
CYTOMETRY
.39
3.2.9
SDS-POLYACRYLAMIDE
GEL
ELECTROPHORESIS
.
40
3.2.10
WESTERN
BLOTTING
.
41
3.2.11
IN
SILICO
MOLECULAR
DOCKING
ANALYSIS
.
42
II
3.2.12
VIABILITY
ASSAY
.
43
3.2.13
F-ACTIN
BINDING
AND
STAINING
.
43
3.2.14
ENDOTHELIAL
CELL
PERMEABILITY
.
44
3.2.15
SCRATCH
ASSAY
.
45
3.2.16
CELLULAR
ROS
DETECTION
.
45
3.2.17
RNA
ISOLATION
.
46
3.2.18
POLYMERASE
CHAIN
REACTION
.
47
3.3
ANIMAL
MODEL
.
51
3.3.1
SURGICAL
IMPLANTATION
OF
OSMOTIC
MINIPUMPS
.52
3.3.2
ATOMIC
FORCE
MICROSCOPY
.
53
3.3.3
TISSUE
FIBROSIS
DETERMINATION
.
54
3.3.4
RNA
ISOLATION
.55
3.3.5
POLYMERASE
CHAIN
REACTION
.
56
3.4
STATISTICAL
ANALYSIS
.
56
3
RESULTS
.
57
3.1
CONFORMATIONAL
CHANGES
OF
THE
ENDOTHELIAL
GLYCOCALYX
UNDER
EXCESS
SFLT-1
.
57
3.1.1
EXCESS
SFLT-1
LEADS
TO
CONFORMATIONAL
CHANGES
OF
THE
EGC
.
57
3.1.2
.
EXCESS
SFLT-1
INCREASES
MONOCYTE
ADHESION
.
62
3.1.3
ENDOTHELIAL
CELL
VIABILITY
UNDER
SFLT-1
.
65
3.2
EXCESS
SFLT-1
AFFECTS
ENDOTHELIAL
STIFFNESS
.
69
3.2.1
EXCESS
SFLT-1
STIFFENS
ENDOTHELIAL
CELL
CORTEX
IN
VITRO
AND
IN
VIVO
.
69
3.2.2
SFLT-1
TREATMENT
ACTIVATES
P38
MAPK
STRESS
RESPONSE
PATHWAY
.
71
III
3.2.3
SFLT-1
INDUCES
CHANGES
IN
THE
F-ACTIN
DISTRIBUTION
AND
DISTURBS
THE
BARRIER
FUNCTION
OF
ENDOTHELIAL
CELLS
.
73
3.2.4
SFLT-1
ONLY
SLIGHTLY
AFFECTS
MECHANOSENSITIVE
GENES
IN
HUVECS
.
76
3.2.5
SFLT-1
AFFECTS
MECHANOSENSITIVE
GENES
IN
THE
MICE
LUNG
.
78
5
DISCUSSION
.
81
5.1
CONFORMATIONAL
CHANGES
OF
THE
ENDOTHELIAL
GLYCOCALYX
UNDER
EXCESS
SFLT-1
.
81
5.2
EXCESS
SFLT-1
AFFECTS
ENDOTHELIAL
STIFFNESS
.
87
5.3
LIMITATIONS
AND
REMARKS
.
92
5.4
CONCLUSION
AND
PERSPECTIVES
.
93
6
APPENDIX
.
94
6.1
MONOCYTE
ADHESION
UNDER
EXCESS
SFLT-1
.
94
6.2
CELL
VIABILITY
UPON
SFLT-1
TREATMENT
.95
6.3
CORTICAL
STIFFNESS
UNDER
TGF-32
TREATMENT
.
96
6.4
P38
MAPK
INHIBITOR
SB203580
PREVENTS
COLLAPSE
OF
THE
EGC
.
97
REFERENCES
.
99
CURRICULUM
VITAE
.119
DANKSAGUNG
.122
IV |
| any_adam_object | 1 |
| any_adam_object_boolean | 1 |
| author | Schulz, Annika 1992- |
| author_GND | (DE-588)1275034012 |
| author_facet | Schulz, Annika 1992- |
| author_role | aut |
| author_sort | Schulz, Annika 1992- |
| author_variant | a s as |
| building | Verbundindex |
| bvnumber | BV048961517 |
| ctrlnum | (OCoLC)1367838916 (DE-599)DNB1275742602 |
| discipline | Medizin |
| discipline_str_mv | Medizin |
| format | Thesis Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV048961517 |
| illustrated | Illustrated |
| index_date | 2024-07-03T22:00:32Z |
| indexdate | 2024-07-10T09:51:21Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-034225268 |
| oclc_num | 1367838916 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR |
| owner_facet | DE-355 DE-BY-UBR |
| physical | XII, 122 Blätter Illustrationen, Diagramme 30 cm |
| publishDate | 2022 |
| publishDateSearch | 2022 |
| publishDateSort | 2022 |
| record_format | marc |
| spelling | Schulz, Annika 1992- Verfasser (DE-588)1275034012 aut Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis vorgelegt von Annika Schulz aus Thuine Münster 2022 XII, 122 Blätter Illustrationen, Diagramme 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Westfälische Wilhelms-Universität Münster 2022 Beschränkt für den Austausch (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1275742602/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034225268&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p emakn 0,04785 20230329 DE-101 https://d-nb.info/provenance/plan#emakn 2\p dnb 20230330 DE-101 https://d-nb.info/provenance/plan#dnb 3\p dnb 20230330 DE-101 https://d-nb.info/provenance/plan#dnb |
| spellingShingle | Schulz, Annika 1992- Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis |
| subject_GND | (DE-588)4113937-9 |
| title | Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis |
| title_auth | Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis |
| title_exact_search | Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis |
| title_exact_search_txtP | Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis |
| title_full | Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis vorgelegt von Annika Schulz aus Thuine |
| title_fullStr | Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis vorgelegt von Annika Schulz aus Thuine |
| title_full_unstemmed | Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis vorgelegt von Annika Schulz aus Thuine |
| title_short | Damage of the endothelial glycocalyx and cortical stiffening - two new mechanisms of action of soluble Fms-like tyrosine kinase (sFlt-1) contributing to endothelial dysfunction beyond angiogenesis |
| title_sort | damage of the endothelial glycocalyx and cortical stiffening two new mechanisms of action of soluble fms like tyrosine kinase sflt 1 contributing to endothelial dysfunction beyond angiogenesis |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1275742602/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=034225268&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT schulzannika damageoftheendothelialglycocalyxandcorticalstiffeningtwonewmechanismsofactionofsolublefmsliketyrosinekinasesflt1contributingtoendothelialdysfunctionbeyondangiogenesis |
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