Verfügbarkeit: Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen

Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen:

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Bibliographische Detailangaben
1. Verfasser:	Schlosser, Monika Stefanie 1985- (VerfasserIn)
Format:	Abschlussarbeit Buch
Sprache:	English
Veröffentlicht:	Köln [2018]
Schlagworte:	Hochschulschrift
Online-Zugang:	Inhaltsverzeichnis Inhaltsverzeichnis
Beschreibung:	119 Seiten Diagramme 21 cm

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Datensatz im Suchindex

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adam_text	1 . TABLE OF CONTENTS 1. TABLE OF CONTENTS 2. GLOSSARY............... 3. INTRODUCTION............................................................................................. 11 3.1 KIDNEY........................................................................................................11 3.1.1 RENAL MORPHOLOGY.............................................................................................................. 11 3.1.2 RENAL PATHOLOGY................................................................................................................. 12 3.2 CELL SIGNALLING PATHWAYS..........................................................................14 3.2.1 MULTIDRUG-RESISTANCE PROTEIN............................................................................................ 14 3.2.2 MITOGEN-ACTIVATED PROTEIN KINASES................................................................................. 14 3.2.3 NUCLEAR FACTOR-KAPPA B .................................................................................................... 15 3.2.4 PROTEIN KINASE C ................................................................................................................ 15 3.2.5 INTERLEUKIN............................................................................................................................15 3.2.6 WNT PATHWAY....................................................................................................................... 15 3.2.7 CYCLOOXYGENASE................................................................................................................. 15 3.2.6 MICRORNAS........................................................................................................................... 16 3.2.9 ENDOCANNABINOID SIGNALLING ............................................................................................ 16 3.2.10 ENDOCANNABINOID RECEPTORS............................................................................................ 16 3.2.11 ENDOTHELIN SIGNALLING..........................................................................................................16 3.2.12 ENDOTHELIN RECEPTOR.......................................................................................................... 20 3.3 CELL DEATH................................................................................................... 22 3.3.1 APOPTOSIS............................................................................................................................ 23 3.3.2 NECROSIS.............................................................................................................................. 24 3.3.3 NECROPTOSIS......................................................................................................................... 24 3.3.4 FERROPTOSIS.................................................................................................. 25 3.4 DISEASE MARKER AND DIAGNOSIS................................................................25 3.5 AIM OF THE STUDY....................................................................................... 26 4. MATERIALS AND METHODS ......................................................................... 27 4.1.1 CHEMICALS........................................................................................................................... 27 4.1.2 BUFFERS AND REAGENTS........................................................................................................ 26 4.1.3 CELL CULTURE......................................................................................................................... 30 4.1.4 CELL LINES............................................................................................................................ 31 4.1.5 PCR REAGENTS................................................................................................................... 31 4.1.6 ANTIBODIES........................................................................................................................... 32 4.1.7 PRIMERS.................................................................................................................................33 4.1.6 EQUIPMENT............................................................................................................................ 33 4.2 METHODS...................................................................................................... 35 4.2.1 CELL CULTURE......................................................................................................................... 35 4.2.2 CELL COUNTING......................................................................................................................35 4.2.3 CELL THAWING AND CELL FREEZING.......................................................................................35 4.2.4 CELL TREATMENT....................................................................................................................36 4.2.5 ADRIAMYCIN MOUSE MODEL OF MURINE PROTEINURIA...........................................................37 4.2.6 BICINCHONINIC ACID ASSAY..................................................................................................38 4.2.7 BRADFORD A SSAY.................................................................................................................. 36 4.2.8 CYTOTOXICITY DETECTION KIT (L D H )......................................................................................36 4.2.9 DETECTION OF REACTIVE OXYGEN SPECIES ......................................................................... 39 4.2.10 ENZYME-IINKED IMMUNOSORBENT ASSAY........... ................................................................. 39 4.2.11 EXTRACTION OF NUCLEAR AND CYTOPLASMIC CELL PROTEIN FRACTION ..................................... 40 4.2.12 EXTRACTION OF RNA AND PURIFICATION.................................................................................41 4.2.13 EXTRACTION OF TOTAL CELL PROTEIN CONTENT..........................................................................41 4.2.14 FACS ANALYSIS................................................................................................................... 42 4.2.15 HISTOLOGY OF PARAFFIN EMBEDDED TISSUE...........................................................................43 4.2.16 HUMAN BIOPSIES................................................................................................................. 43 4.2.17 HPLC-MS/MS ANALYSIS.....................................................................................................43 4.2.16 IMMUNOPRECIPITATION ASSAY (IP )....................................................................................... 45 IO H- 4.2.19 LC/MRM ANALYSIS.............................................................................................................. 45 4.2.20 MIRNA ISOLATION.................................................................................................................45 4.2.21 NEPHRECTOMY ANIMAL MODEL.............................................................................................46 4.2.22 POLYMERASE CHAIN REACTION............................................................................................46 4.2.23 QUANTITATIVE REAL-TIME POLYMERASE CHAIN REACTION...................................................47 4.2.24 RENAL TRANSPLANT MODEL OF ACUTE RENAL INJURY.............................................................47 4.2.25 REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION....................................................48 4.2.26 RNA QUANTIFICATION........................................................................................................... 48 4.2.27 WESTERN BLOT....................................................................................................................... 49 4.2.26 STATISTICS............................................................................................................................... 50 5. R ESULTS.....................................................................................................51 5.1 RENAL CELLS EXPRESS ENDOCANNABINOID SYSTEM....................................51 5.1.1 DETERMINATION OF LIPIDS 2-AG AND AEA IN KIDNEY SAMPLES USING LIQUID CHROMATOGRAPHY...............................................................................................................51 5.1.2 DETERMINATION OF ENDOCANNABINOID RECEPTORS AND FAAH IN RENAL CELLS ON RNA LEVEL....................................................................................................................................52 5.1.3 HISTOLOGY OF NAPE-PLD AND FAAH IN HUMAN RENAL TISSUE......................................54 5.1.4 DETERMINATION OF ENDOCANNABINOID RECEPTORS IN RENAL CELLS AT PROTEIN LEVEL ...... 55 5.2 INCREASED CELL DEATH OF RENAL PROXIMAL TUBULE CELLS UPON STIMULATION WITH AEA...................................................................................................57 5.2.1 LDH RELEASE AFTER AEA TREATMENT OF RPTEC AND CAKI-1 ........................................ 57 5.2.2 MICROSCOPIC MORPHOLOGY UPON AEA TREATMENT............................................................58 5.2.3 RENAL PROXIMAL TUBULE CELL DEATH BY AEA IS NOT MEDIATED VIA ENDOCANNABINOID RECEPTORS............................................................................................................................59 5.2.4 RENAL PROXIMAL TUBULE CELL DEATH BY AEA IS MEDIATED VIA R O S ............................. 60 5.2.5 STRONG EVIDENCE FOR FERROPTOSIS AS MAIN COMPONENT OF TUBULAER CELL DEATH IN CAKI- 1 AND RPTEC INITIALISED BY AEA..................................................................................... 63 5.3 INTRACELLULAR ENDOCANNABINOID SYSTEM AND ENDOTHELIN SYSTEM INDICATE OVERLAP OF SIGNALLING PATHWAYS ON RNA AND PROTEIN LEVEL..64 5.3.1 SEQUENCE ALIGNMENTS OF ENDOTHELIN RECEPTORS AND CANNABINOID RECEPTORS ........ 64 5.3.2 INTERFERENCE OF ENDOCANNABINOID SYSTEM AND ENDOTHELIN SYSTEM ...........................66 5.3.3 ENDOCANNABINOID AND ENDOTHELIN SIGNALLING IN RATS WITH ETBR KNOCKOUT ............ 69 5.4 CLINICAL RELEVANCE.....................................................................................71 5.4.1 CONNECTION BETWEEN ENDOTHELIN SIGNALLING AND M R P2...............................................71 5.4.2 MRP2 IN RENAL TISSUE OF MEMBRANOUS NEPHROPATHY, MINIMAL CHANGE NEPHROPATHY AND ADRIAMYCIN MOUSE MODEL ................................................................. 72 5.4.3 MIRNA133A EXPRESSION IN URI NE OF MEMBRANOUS NEPHROPATHY, MINIMAL CHANGE NEPHROPATHY AND ADRIAMYCIN MOUSE MODEL ................................................................. 74 6. DISCUSSION..............................................................................................76 6.1 GENERAL REMARKS ON ENDOCANNABINOIDS AND CELL DEATH ..................... 76 6.2 RENAL CELLS EXPRESS ENDOCANNABINOID SYSTEM....................................79 6.3 INCREASED CELL DEATH OF RENAL PROXIMAL TUBULE CELLS UPON STIMULATION WITH AEA...................................................................................................85 6.4 INTRACELLULAR ENDOCANNABINOID SYSTEM AND ENDOTHELIN SYSTEM INDICATE OVERLAP OF SIGNALLING PATHWAYS ON RNA AND PROTEIN LEVEL..89 6.5 CONNECTION BETWEEN ENDOTHELIN SIGNALLING AND MRP2 ....................... 93 6.6 CLINICAL RELEVANCE, SUMMARY AND CONCLUSION......................................95 7. SUM MARY................................................................................................. 99 8. ZUSAMMENFASSUNG..............................................................................100 9. REFERENCES............................................................................................101 10. PRELIMINARY PUBLICATIONS....................................................................116 11. LEBENSLAUF............................................................................................118
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spelling	Schlosser, Monika Stefanie 1985- Verfasser (DE-588)1165252295 aut Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen vorgelegt von Monika Stefanie Schlosser Köln [2018] 119 Seiten Diagramme 21 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Universität zu Köln 2018 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf http://d-nb.info/1165808579/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031280681&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis
spellingShingle	Schlosser, Monika Stefanie 1985- Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen
subject_GND	(DE-588)4113937-9
title	Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen
title_auth	Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen
title_exact_search	Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen
title_full	Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen vorgelegt von Monika Stefanie Schlosser
title_fullStr	Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen vorgelegt von Monika Stefanie Schlosser
title_full_unstemmed	Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen vorgelegt von Monika Stefanie Schlosser
title_short	Synergistischer Effekt von Endothelin-1 Signalling und dem Endocannabinoid System in Renalen Proximalen Tubulus Zellen
title_sort	synergistischer effekt von endothelin 1 signalling und dem endocannabinoid system in renalen proximalen tubulus zellen
topic_facet	Hochschulschrift
url	http://d-nb.info/1165808579/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031280681&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA
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