Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide: insights into mechanisms and mode of cell death
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
2002
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | München, Univ., Fak. für Chemie und Pharmazie, Diss., 2002 |
| Beschreibung: | [5], 113 S. Ill., graph. Darst. : 21 cm |
Internformat
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV014767952 | ||
| 003 | DE-604 | ||
| 005 | 20021203 | ||
| 007 | t | ||
| 008 | 020924s2002 gw ad|| m||| 00||| eng d | ||
| 016 | 7 | |a 965030067 |2 DE-101 | |
| 035 | |a (OCoLC)52653602 | ||
| 035 | |a (DE-599)BVBBV014767952 | ||
| 040 | |a DE-604 |b ger |e rakddb | ||
| 041 | 0 | |a eng | |
| 044 | |a gw |c DE | ||
| 049 | |a DE-12 |a DE-19 | ||
| 100 | 1 | |a Gerwig, Tobias |d 1972- |e Verfasser |0 (DE-588)124004539 |4 aut | |
| 245 | 1 | 0 | |a Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide |b insights into mechanisms and mode of cell death |c von Tobias Gerwig |
| 264 | 1 | |c 2002 | |
| 300 | |a [5], 113 S. |b Ill., graph. Darst. : 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a München, Univ., Fak. für Chemie und Pharmazie, Diss., 2002 | ||
| 650 | 7 | |a Hochschulschrift |2 gtt | |
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 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=009999853&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-009999853 | |
Datensatz im Suchindex
| _version_ | 1807593335361110016 |
|---|---|
| adam_text |
CONTENTS
CONTENTS
A.
INTRODUCTION
.
1
1.
BACKGROUND
AND
OVERVIEW
.
1
2.
ISCHEMIA-REPERFUSION
INJURY
OF
THE
LIVER
.
3
2.1
SIGNIFICANCE
.
3
2.2
PATHOMECHANISMS
IN
COLD
ISCHEMIA
AND
REPERFUSION
.
4
2.2.1
ISCHEMIA
.
4
2.2.2
REPERFUSION
.
4
3.
MODE
OF
CELL
DEATH
.
6
3.1
NECROSIS
.
6
3.2
APOPTOSIS
.
7
3.2.1
MECHANISMS
.
8
4.
THERAPEUTIC
STRATEGIES
.
11
4.1
ESTABLISHED
THERAPIES
.
11
4.1.1
STORAGE
AND
RINSE
SOLUTIONS
.
11
4.1.2
PROSTAGLANDINS
.
11
4.2
NEW
THERAPEUTIC
ADVANCES
.
12
4.2.1
ISCHEMIC
PRECONDITIONING
.
12
4.2.2
ANTIOXIDATIVE
STRATEGIES
.
12
4.2.3
HORMONAL
PRECONDITIONING
WITH
THE
ATRIAL
NATRIURETIC
PEPTIDE
.
12
5.
THE
ATRIAL
NATRIURETIC
PEPTIDE
.
14
5.1
NATRIURETIC
PEPTIDES
.
14
5.2
RECEPTORS
AND
SIGNAL
TRANSDUCTION
.
15
5.3
CGMP-DEPENDENT
SIGNALING
.
16
5.4
CYCLIC
NUCLEOTIDE
DEPENDENT
PROTEIN
KINASES
.
17
5.4.1
CGMP-DEPENDENT
PROTEIN
KINASES
.
18
5.4.2
CAMP-DEPENDENT
PROTEIN
KINASES
.
19
6.
THE
HEME
OXYGENASE
SYSTEM
.
20
6.1
BIOLOGICAL
FUNCTIONS
.
20
6.2
HEME
OXYGENASE-1
IN
OXIDANT
STRESS
.
21
6.3
HEME
OXYGENASE-1
:
SIGNIFICANCE
IN
IRI
.
21
7.
THE
ISOLATED
PERFUSED
RAT
LIVER:
GENERAL
ASPECTS
.
22
B.
MATERIALS
AND
METHODS
.
23
1.
THE
ISOLATED
PERFUSED
RAT
LIVER
.
23
CONTENTS
1.1
ANIMALS
.
23
1.2
SOLUTIONS
.
23
1.3
MODEL
OF
THE
ISOLATED
PERFUSED
RAT
LIVER
.
23
1.4
EXPERIMENTAL
SETTING
.
25
1.4.1
CONTINOUS
LIVER
PERFUSION
.
25
1.4.2
MODEL
OF
COLD
ISCHEMIA
AND
REPERFUSION
.
25
1.4.3
TREATMENT
PROTOCOLS
.
25
1.5
PARAMETERS
OF
LIVER
FUNCTION
.
26
1.5.1
LACTATE
DEHYDROGENASE
EFFLUX
.
26
1.5.2
BILE
EFFLUX
.
26
1.5.3
PORTAL
PRESSURE
.
27
2.
LIVER
CELL
CULTURE
.
27
2.1
ANIMALS
.
27
2.2
SOLUTIONS
.
27
2.3
CULTIVATION
.
28
2.4
ISOLATION
OF
PRIMARY
HEPATOCYTES
.
29
2.5
ISOLATION
OF
KUPFFER
CELLS
.
30
3.
ISOLATION
OF
RIBONUCLEIC
ACIDS
.
31
3.1
EXTRACTION
OF
RIBONUCLEIC
ADDS
.
31
3.1.1
SOLUTIONS
.
31
3.1.2
GUANIDINE
THIOCYANATE/CESIUMCHLORIDE-METHOD
.
31
3.2
MEASUREMENT
OF
NUDEIC
ADD
CONCENTRATION
.
32
3.3
GEL
ELECTROPHORETIC
SEPARATION
OF
NUCLEIC
ACIDS
.
32
3.3.1
SOLUTIONS
.
33
3.3.2
AGAROSE
GEL
ELECTROPHORESIS
.
33
3.3.3
STAINING
.
33
3.4
SEPARATION
OF
MRNA
FROM
TOTRNA
.
34
4.
NORTHERN
BLOT
.
34
4.1
BADERIA
.
34
4.1.1
STRAIN
.
34
4.1.2
GROWTH
MEDIA
AND
ANTIBIOTICS
.
35
4.1.3
CULTIVATION
.
35
4.2
VECTORS
.
35
4.2.1
EMPLOYED
PLASMIDS
.
35
4.3
AMPLIFICATION
OF
A
CDNA-NORTHEM
BLOT
PROBE
.
36
4.3.1
PREPARATION
OF
COMPETENT
CELLS
.
36
4.3.2
TRANSFORMATION
OF
COMPETENT
E.
COLL
.
36
CONTENTS
4.3.3
MINI-PREP
DNA
ISOLATION
.
37
4.3.4
AGAROSE
GEL
ELECTROPHORESIS
.
37
4.3.5
MIDI-PREP
DNA
ISOLATION
WITH
QIAGEN
PLASMID
MIDI
KIT
.
38
4.3.6
RESTRICTION
ENDONUCLEASE
DIGESTION
.
39
4.3.7
EXTRACTION
OF
DNA
FROM
AGAROSE
GELS
.
39
4.3.8
SEQUENCING
OF
DOUBLE
STRANDED
DNA
.
40
4.4
DNA
LABELING
WITH
RADIOACTIVE
NUCLEOTIDES
.
40
4.4.1
SYNTHESIS
OF
RADIOACTIVELY
LABELED
HO-1
CDNA
.
40
4.4.2
SYNTHESIS
OF
RADIOACTIVELY
LABELED
28S
RNA
OLIGONUCLEOTIDES
.
41
4.4.3
REMOVAL
OF
UNINCORPORATED
DNTPS
.
41
4.5
NORTHERN
BLOT
.
42
4.5.1
SOLUTIONS
.
42
4.5.2
TRANSFER
OF
ELECTROPHORETICALLY
SEPARATED
RNA
ONTO
A
NYLON
MEMBRANE
.
42
4.5.3
HYBRIDIZATION
.
43
4.5.4
DETECTION
.
43
5.
IN
SITU
HYBRIDIZATION
.
43
5.1
HYBRIDIZATION
.
43
5.2
DETECTION
.
44
6.
REVERSE
TRANSCRIPTASE
POLYMERASE
CHAIN
REACTION
.
44
6.1
REVERSE
TRANSCRIPTION
.
44
6.2
PRIMERS
AND
REACTION
CONDITIONS
.
45
6.3
POLYMERASE
CHAIN
REACTION
.
45
6.4
ELECTROPHORESIS
OF
AMPLIFICATION
PRODUCTS
.
46
7.
DETECTION
OF
PROTEINS
BY
WESTERN
BLOTTING
.
46
7.1
PREPARATION
OF
SAMPLES
.
46
7.1.1
SOLUTIONS
.
46
7.1.2
PREPARATION
OF
CELLULAR
PROTEIN
EXTRACTS
FROM
RAT
LIVER
TISSUE
.
47
7.1.3
PREPARATION
OF
CELLULAR
PROTEIN
EXTRACTS
FROM
RAT
HEPATOCYTES
.
47
7.1.4
PREPARATION
OF
SAMPLES
FOR
PROTEIN
ELECTROPHORESIS
.
47
7.2
MEASUREMENT
OF
PROTEIN
CONCENTRATION:
LOWRY-ASSAY
.
47
7.3
SDS-POLYACRYLAMIDE
GEL
ELECTROPHORESIS
(SDS-PAGE)
.
48
7.3.1
SOLUTIONS
.
48
7.3.2
ELECTROPHORESIS
.
49
7.4
COOMASSIE
BLUE
STAINING
.
49
7.4.1
SOLUTIONS
.
49
7.4.2
STAINING
PROCEDURE
.
50
7.5
IMMUNOLOGIC
DETECTION
BY
WESTERN
BLOTTING
.
50
CONTENTS
7.5.1
SOLUTIONS
.
50
7.5.2
PROTEIN
TRANSFER
BY
SEMIDRY
BLOTTING
.
51
7.5.3
USED
ANTIBODIES
.
51
7.5.4
INCUBATION
WITH
ANTIBODIES
AND
CHEMILUMINESCENCE
DETECTION
.
52
8.
IN
VITRO
PHOSPHORYLATION
BY
PROTEIN
KINASE
A
.
52
8.1
SOLUTIONS
.
52
8.2
PREPARATION
OF
SAMPLES
.
53
8.2.1
LIVER
TISSUE
.
53
8.2.2
PRIMARY
HEPATOCYTES
.
53
8.3
PROTEIN
KINASE
A
ASSAY
.
53
9.
FLUORIMETRIC
ENZYME
ACTIVITY
ASSAY
.54
9.1
SOLUTIONS
.
55
9.2
CASPASE-3-LIKE
ADTIVITY
ASSAY
.
55
9.3
PROTEIN
DETERMINATION:
PIERCE-ASSAY
(BCA-ASSAY)
.
55
10.
HISTOLOGICAL
ANALYSIS
.
56
10.1
TUNEL
STAINING
.
56
10.2
IMMUNOHISTOCHEMISTRY
.
56
10.3
HEMATOXYLIN/EOSIN
(H/E)
STAINING
.
57
10.4
TRYPAN
BLUE
UPTAKE
OF
LIVER
TISSUE
.
57
11.
STATISTICS
.
58
C.
RESULTS
.
59
1.
DETERMINATION
OF
APOPTOTIC
AND
NECROTIC
CELL
DEATH
.
59
1.1
EVALUATION
OF
APOPTOSIS
.
59
1.1.1
ANP
AND
8-BR-CGMP
PRECONDITIONING
REDUCE
CASPASE-3-LIKE
ACTIVITY
AND
CASPASE-3
PROCESSING
.
59
1.1.2
ANP
AND
8-BR-CGMP
PRECONDITIONING
DECREASE
THE
PROPORTION
OF
TUNEL
POSITIVE
CELLS
.
63
1.2
EVALUATION
OF
NECROSIS
.
64
1.2.1
DECREASED
DEGENERATIVE
CHANGES
IN
ANP
PRE-TREATED
LIVERS
.
64
1.2.2
TRYPAN
BLUE
UPTAKE
IS
SIGNIFICANTLY
REDUCED
BY
ANP
PRECONDITIONING
.
65
2.
SIGNAL
TRANSDUCTION
OF
ANP
PROTECTION
.
66
2.1
INVOLVEMENT
OF
CGMP-DEPENDENT
PROTEIN
KINASES
.
66
2.2
EFFECT
OF
CGMP
AND
CAMP-DEPENDENT
PROTEIN
KINASE
INHIBITORS
ON
THE
ANTIAPOPTOTIC
PROPERTY
OF
ANP
.
68
2.2.1
EFFECTS
ON
CASPASE-3-LIKE
ACTIVITY
.
68
2.3
ACTIVATION
OF
CAMP-DEPENDENT
PROTEIN
KINASE
BY
ANP
.
69
CONTENTS
2.3.1
LIVER
.
69
2.3.2
HEPATOCYTES
.
70
3.
HEME
OXYGENASE-1
.
71
3.1
CGMP-INDEPENDENT
INDUCTION
OF
HO-1
BY
ANP
PRECONDITIONING
.
71
3.2
LOCALIZATION
OF
HO-1
.
74
3.3
INDUCTION
OF
HO-1
IN
ISOLATED
KUPFFER
CELLS
.
75
3.4
ZNPP
PERFUSION
.
76
D.
DISCUSSION
.
78
1.
REDUCTION
OF
APOPTOSIS
AND
NECROSIS
BY
ANP
.
78
1.1
REDUCTION
OF
APOPTOSIS
BY
ANP
TREATMENT
.
78
1.2
CASPASE-3
UPREGULATION
DURING
COLD
ISCHEMIA
.
79
1.3
ANP
REDUCES
EARLY
DEGENERATIVE
CELL
CHANGES
.
79
1.4
REDUCTION
OF
NECROSIS
BY
ANP
TREATMENT
.
80
2.
SIGNAL
TRANSDUCTION
OF
ANP
PROTECTION
.
80
2.1
CGM
P-DEPENDENT
PROTEIN
KINASES
.
80
2.2
CAMP-DEPENDENT
PROTEIN
KINASES
.
81
2.3
FURTHER
POTENTIALLY
INVOLVED
PATHWAYS
.
82
3.
HEME
OXYGENASE-1
EXPRESSION
.
83
3.1
HO-1
EXPRESSION
DURING
ISCHEMIA
AND
REPERFUSION
.
83
3.2
ANP
INDUCES
HO-1
IN
RAT
LIVERS
.
83
3.3
INDUCTION
OF
HO-1
IN
KUPFFER
CELLS
.
84
3.4
INDUCTION
OF
HO-1
BY
ANP
IS
INDEPENDENT
OF
CGMP
.
84
3.5
PERFUSION
WITH
ZNPP
.
85
E.
SUMMARY
.86
F.
APPENDIX
.
87
1.
ABBREVIATIONS
.
87
2.
ALPHABETICAL
ORDER
OF
COMPANIES
.
90
3.
PUBLICATIONS
.92
3.1
ABSTRACTS
.
92
3.2
ORIGINAL
PUBLICATIONS
.
93
G.
REFERENCES
.
94
H.
ACKNOWLEGEMENTS
.
112
I.
CURRICULUM
VITAE
.
113 |
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| institution | BVB |
| language | English |
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| spelling | Gerwig, Tobias 1972- Verfasser (DE-588)124004539 aut Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide insights into mechanisms and mode of cell death von Tobias Gerwig 2002 [5], 113 S. Ill., graph. Darst. : 21 cm txt rdacontent n rdamedia nc rdacarrier München, Univ., Fak. für Chemie und Pharmazie, Diss., 2002 Hochschulschrift gtt (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009999853&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Gerwig, Tobias 1972- Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide insights into mechanisms and mode of cell death Hochschulschrift gtt |
| subject_GND | (DE-588)4113937-9 |
| title | Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide insights into mechanisms and mode of cell death |
| title_auth | Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide insights into mechanisms and mode of cell death |
| title_exact_search | Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide insights into mechanisms and mode of cell death |
| title_full | Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide insights into mechanisms and mode of cell death von Tobias Gerwig |
| title_fullStr | Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide insights into mechanisms and mode of cell death von Tobias Gerwig |
| title_full_unstemmed | Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide insights into mechanisms and mode of cell death von Tobias Gerwig |
| title_short | Prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide |
| title_sort | prevention of ischemia reperfusion injury in the rat liver by atrial natriuretic peptide insights into mechanisms and mode of cell death |
| title_sub | insights into mechanisms and mode of cell death |
| topic | Hochschulschrift gtt |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009999853&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT gerwigtobias preventionofischemiareperfusioninjuryintheratliverbyatrialnatriureticpeptideinsightsintomechanismsandmodeofcelldeath |