Analysis of the effects of xanthohumol on hepatic homeostasis, inflammation, fibrosis and cancerogenesis [Elektronische Ressource] / vorgelegt von Christoph Michael Dorn

ANALYSIS OF THE EFFECTS OF XANTHOHUMOL
ON HEPATIC HOMEOSTASIS, INFLAMMATION, FIBROSIS
AND CANCEROGENESIS







DISSERTATION

ZUR ERLANGUNG DES DOKTORGRADES DER NATURWISSENSCHAFTEN
(DR. RER. NAT.) DER FAKULTÄT CHEMIE UND PHARMAZIE
DER UNIVERSITÄT REGENSBURG







vorgelegt von
Christoph Michael Dorn
aus Höchstädt a.d. Donau
im Jahr 2009
























Promotionsgesuch eingereicht: Januar 2007

Die Arbeit wurde angeleitet von: Herrn PD Dr. Claus Hellerbrand

Prüfungsausschuss:
Vorsitzender: Herr Prof. Dr. Elz
1. Gutachter (1. Prüfer): Herr Prof. Dr. Jörg Heilmann
2. Gutachter (2. Prüfer): Herr PD Dr. Claus Hellerbrand
3. Prüfer: Herr Prof. Dr. Schlossmann




























für meine Familie





I
Table of Contents
1 SUMMARY ................................................................................................................................ 1
2 INTRODUCTION ..................................................................................................................... 3
2.1 HOP........................................................................................................................................... 3
2.1.1 BOTANY ................................................................................................................................. 3
2.1.2 MEDICAL USE......................................................................................................................... 4
2.1.3 PHYTOCHEMISTRY ................................................................................................................. 5
2.2 FLAVONOIDS ............................................................................................................................ 6
2.3 XANTHOHUMOL....................................................................................................................... 8
2.3.1 DIETARY EXPOSURE............................................................................................................... 8
2.3.2 BIOAVAILABILITY AND METABOLISM.................................................................................... 9
2.4 BIOLOGICAL EFFECTS OF XANTHOHUMOL.......................................................................... 11
2.4.1 ANTI-INFLAMMATORY EFFECTS........................................................................................... 11
2.4.2 ANTI-ANGIOGENIC EFFECTS................................................................................................. 12
2.4.3 ANTI-CANCEROUS EFFECTS.................................................................................................. 14
2.4.4 ANTIOXIDANT PROPERTIES .................................................................................................. 17
2.4.5 ANTI-INFECTIVE PROPERTIES............................................................................................... 18
2.4.5.1 Anti-bacterial effects......................................................................................................... 18
2.4.5.2 Anti-fungal effects ............................................................................................................ 19
2.4.5.3 Anti-malarial effects ......................................................................................................... 19
2.4.5.4 Anti-viral effects............................................................................................................... 20
2.4.6 (ANTI)ESTROGENIC POTENTIAL ........................................................................................... 21
2.4.7 EFFECTS ON LIPID AND CARBOHYDRATE METABOLISM....................................................... 22
2.4.8 EFFECTS ON BONE RESORPTION ........................................................................................... 23
2.5 XANTHOHUMOL SAFETY STUDIES ........................................................................................ 23
2.6 EFFECTS OF XANTHOHUMOL ON THE LIVER ....................................................................... 25
2.6.1 EFFECTS OF ORAL ADMINISTERED XANTHOHUMOL ON THE LIVER...................................... 26
2.6.2 EFFECTS OF XANTHOHUMOL ON HEPATOCELLULAR CARCINOMA CELLS ............................ 26
2.6.3 EFFECTS OF XANTHOHUMOL ON HEPATOCYTES .................................................................. 27
2.6.4 EFFECTS OF XANTHOHUMOL ON NON-PARENCHYMAL LIVER CELLS ................................... 27
2.7 LIVER DISEASES..................................................................................................................... 28
2.7.1 DEFINITION AND NATURAL COURSE OF LIVER DISEASE....................................................... 28
2.7.2 LIVER FIBROSIS .................................................................................................................... 28
2.7.3 LIVER CIRRHOSIS ................................................................................................................. 30 Table of Contents II
2.7.4 LIVER CANCER ..................................................................................................................... 31
2.7.4.1 Hepatocellular cancer........................................................................................................ 31
2.7.4.1.1 Prevalence and incidence............................................................................................... 31
2.7.4.1.2 Etiology.......................................................................................................................... 31
2.7.4.1.3 Therapy and prognosis................................................................................................... 32
2.7.5 CAUSES FOR CHRONIC LIVER DISEASES ............................................................................... 32
2.7.5.1 Alcohol induced liver disease ........................................................................................... 33
2.7.5.2 Drug induced liver injury.................................................................................................. 34
2.7.5.3 Autoimmune related mechanisms and genetic defects ..................................................... 34
2.7.5.4 Viral hepatitis.................................................................................................................... 35
2.7.5.4.1 Hepatitis A ..................................................................................................................... 35
2.7.5.4.2 Hepatitis B ..................................................................................................................... 35
2.7.5.4.3 Hepatitis C ..................................................................................................................... 36
2.7.5.5 Non-alcoholic fatty liver disease (NAFLD)...................................................................... 36
2.7.5.5.1 Definition....................................................................................................................... 36
2.7.5.5.2 Prevalence of NAFLD/NASH ....................................................................................... 37
2.7.5.5.3 Etiology and pathogenesis ............................................................................................. 38
2.7.5.5.4 Prognosis and therapy.................................................................................................... 38
2.7.5.5.5 Experimental NASH models.......................................................................................... 39
2.8 AIM OF THE THESIS ............................................................................................................... 40
3 MATERIALS AND METHODS ............................................................................................ 41
3.1 CHEMICALS AND REAGENTS................................................................................................. 41
3.2 LABORATORY EXPENDABLES................................................................................................ 41
3.3 LABORATORY INSTRUMENTS................................................................................................ 42
3.4 BUFFERS................................................................................................................................. 43
3.5 CELL CULTURE...................................................................................................................... 43
3.5.1 CELL CULTURE MEDIUM....................................................................................................... 43
3.5.2 CULTIVATION OF CELL LINES............................................................................................... 44
3.5.3 HUMAN HEPATOCELLULAR CARCINOMA CELL LINES .......................................................... 44
3.5.4 ISOLATION OF PRIMARY HUMAN HEPATOCYTES.................................................................. 44
3.5.5