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Kristýna Paclíková Influence of resveratrol on bile flow Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Pharmacy Background: Protective effect of resveratrol is shown in many experimental models of cholestasis, but its effect on bile production in healthy individuals has not been studied yet. As the use of resveratrol is frequent in population, the aim of this thesis was to investigate the effect of resveratrol on the bile flow in healthy animals and the clarification of the mechanism of this effect. Methods: Wistar rats (n = 6, in each group, weighing 280 to 320 g) were divided into two groups: control group (Control) and the group of rats administered with resveratrol (10mg/kg/day, p.o.) for 28 days (RES). In vivo clearance study was performed to analyze bile production. Analysis of mRNA and protein expression of the transport proteins was performed by qRT-PCR and Western blot. Results: Resveratrol led to a significant increase in the cumulative bile flow. Analysis of mRNA and protein expression of the transport proteins revealed that bile flow is changed due to posttranscriptional induction of canalicular transporters Bsep and Mrp2. Simultaneously, resveratrol led to induction of Mrp4 efflux protein, expressed at the basolateral membrane of hepatocytes....

Wong Hei Yi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 157-175). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Abstract 摘要 --- p.iv / List of Figures --- p.vi / List of Tables --- p.ix / List of Abbreviations --- p.x / Table of Contents --- p.xiii / Chapter Chapter 1: --- Introduction --- p.1 / Chapter 1.1 --- Importance of Glucose in Biological System --- p.1 / Chapter 1.2 --- Glucose Transporter Families --- p.2 / Chapter 1.2.1 --- Na+-Dependent Glucose Transporters --- p.2 / Chapter 1.2.2 --- Facilitative Glucose Transporters --- p.3 / Chapter 1.3 --- Glucose Transporter Type1 --- p.7 / Chapter 1.3.1 --- Primary Structure --- p.7 / Chapter 1.3.2 --- Secondary Structure --- p.8 / Chapter 1.3.3 --- Membrane Topology --- p.8 / Chapter 1.3.4 --- Tertiary Structure --- p.9 / Chapter 1.3.5 --- Kinetics Properties --- p.11 / Chapter 1.3.6 --- Affinity Reagents --- p.12 / Chapter 1.3.7 --- Tissue Distribution --- p.13 / Chapter 1.3.8 --- Multifunctional Property --- p.14 / Chapter 1.3.9 --- Characterization of GLUT1 Gene --- p.14 / Chapter 1.3.10 --- Regulation of GLUT1 Expression --- p.15 / Chapter 1.4 --- Glucose Transporter Type 1 and the Brain --- p.17 / Chapter 1.5 --- Glucose Transporter Type 1 Deficiency Syndrome --- p.20 / Chapter 1.5.1 --- Background of GlutlDS --- p.20 / Chapter 1.5.2 --- Clinical Features of GlutlDS --- p.23 / Chapter 1.5.3 --- Genotype-Phenotype Correlations --- p.24 / Chapter 1.5.4 --- Diagnosis --- p.26 / Chapter 1.5.4.1 --- Erythrocyte Glucose Transporter Activity --- p.26 / Chapter 1.5.4.2 --- Molecular Genetic Testing of GLUT1 Gene --- p.27 / Chapter 1.5.4.3 --- Glucose Concentration --- p.27 / Chapter 1.5.5 --- Management --- p.28 / Chapter 1.5.5.1 --- Ketogenic Diet --- p.28 / Chapter 1.5.5.2 --- Medication --- p.29 / Chapter 1.5.5.2.1 --- Glutl Activator --- p.29 / Chapter 1.5.5.2.2 --- Glutl Inhibitor --- p.29 / Chapter 1.6 --- Hypothesis and Objectives --- p.31 / Chapter Chapter 2: --- Identification of the First Two Asian GlutlDS Cases --- p.33 / Chapter 2.1 --- Materials --- p.34 / Chapter 2.1.1 --- Clinical History of Suspected GlutlDS Patients --- p.34 / Chapter 2.1.2 --- Blood Samples --- p.35 / Chapter 2.1.3 --- Reagents for Zero-trans Influx of 3-OMG Uptake in Erythrocytes --- p.35 / Chapter 2.1.4 --- Reagents for Zero-trans Efflux of 3-OMG Uptake in Erythrocytes --- p.37 / Chapter 2.1.5 --- Reagents for Glutl Gene Analysis --- p.37 / Chapter 2.1.6 --- Reagents and Buffers for Reverse Transcription --- p.38 / Chapter 2.1.7 --- Reagents and Buffers for Agarose Gel Electrophoresis --- p.39 / Chapter 2.1.8 --- Reagents for Erythrocytes Membrane Preparation and Detection --- p.41 / Chapter 2.2 --- Methods --- p.46 / Chapter 2.2.1 --- Zero-trans Influx of 3-OMG Uptake in Erythrocytes --- p.46 / Chapter 2.2.2 --- Zero-trans Efflux of 3-OMG out of Erythrocytes --- p.47 / Chapter 2.2.3 --- Glutl Protein Expression --- p.48 / Chapter 2.2.4 --- GLUT1 Gene Analyses --- p.51 / Chapter 2.2.5 --- Statistics --- p.58 / Chapter 2.3 --- Results --- p.59 / Chapter 2.4 --- Discussions and Conclusions --- p.69 / Chapter Chapter 3: --- Pathogenicity of GLUT1 Mutations --- p.78 / Chapter 3.1 --- Materials --- p.79 / Chapter 3.1.1 --- Construction of Glutl-Encoding Vectors --- p.79 / Chapter 3.1.2 --- Cell Lines --- p.80 / Chapter 3.1.3 --- "Cell Culture Media, Buffers and Other Reagents" --- p.81 / Chapter 3.1.4 --- Cell Culture Wares --- p.83 / Chapter 3.1.5 --- Reagents for Transfection --- p.83 / Chapter 3.1.6 --- Reagents for Protein Determination and Western Blot Analysis --- p.83 / Chapter 3.1.7 --- Reagents and Buffers for Flow Cytometry --- p.84 / Chapter 3.1.8 --- Reagents for 2-DOG Uptake in CHO-K1 Cells --- p.84 / Chapter 3.1.9 --- Reagents and Consumables for Confocal Microscopy --- p.85 / Chapter 3.2 --- Methods --- p.86 / Chapter 3.2.1 --- Cell Culture Methodology --- p.86 / Chapter 3.2.2 --- Construction of Glutl-Encoding Vectors --- p.87 / Chapter 3.2.3 --- Construction of Glutl Mutants --- p.91 / Chapter 3.2.4 --- Establishment of Wild Type and Mutant Glutl Expressing Cell Lines --- p.92 / Chapter 3.2.5 --- Glucose Influx Assays in CHO-K1 Cells --- p.96 / Chapter 3.2.6 --- Confocal Microscopy Studies on Glutl Cellular Localization --- p.97 / Chapter 3.2.7 --- Statistics --- p.98 / Chapter 3.3 --- Results --- p.99 / Chapter 3.4 --- Discussions and Conclusions --- p.112 / Chapter Chapter 4: --- Effects of Anticonvulsive Compounds on Cellular Glucose Transport --- p.117 / Chapter 4.1 --- Materials --- p.118 / Chapter 4.1.1 --- Cell Lines --- p.118 / Chapter 4.1.2 --- Cell Culture Media --- p.118 / Chapter 4.1.3 --- Blood Sample --- p.119 / Chapter 4.1.4 --- Anticonvulsive Compounds --- p.119 / Chapter 4.1.5 --- Reagents for Zero-trans Influx of 3-OMG Uptake in Fibroblasts --- p.120 / Chapter 4.1.6 --- Reagents for Zero-trans Influx of 2-DOG Uptake in Primary Astrocytes --- p.120 / Chapter 4.1.7 --- Reagents for Total RNA Isolation --- p.121 / Chapter 4.1.8 --- Reagents and Consumables for Real-Time PCR --- p.122 / Chapter 4.2 --- Methods --- p.123 / Chapter 4.2.1 --- Cell Culture --- p.123 / Chapter 4.2.2 --- Drug Concentrations --- p.123 / Chapter 4.2.3 --- Zero-trans Influx of 3-OMG Uptake in Erythrocytes --- p.123 / Chapter 4.2.4 --- Zero-trans Influx of 3-OMG Uptake in Fibroblasts --- p.124 / Chapter 4.2.5 --- Zero-trans Influx of 2-DOG Uptake in Primary Astrocytes --- p.125 / Chapter 4.2.6 --- Gene Expression Study --- p.127 / Chapter 4.2.7 --- Statistics --- p.130 / Chapter 4.3 --- Results --- p.131 / Chapter 4.4 --- Discussions and Conclusions --- p.148 / Chapter Chapter 5: --- General Conclusions and Future Perspectives --- p.154 / References --- p.157

Diss. (sammanfattning) Uppsala : Univ., 2003. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Universitet, 2005. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 7 uppsatser.

Thesis (Ph. D.)--University of Washington, 1996. / Vita. Includes bibliographical references (p. [167]-180).