Effects of tumor necrosis factor-alpha on glucose uptake in primary cultured rat astrocytes.

• Other Authors: Wong, Chun Lung.
• Format: Others
• Language: English; Chinese
• Published: 2005
• Subjects: Tumor necrosis factor; Glucose > Metabolism; Astrocytes; Tumor Necrosis Factor-alpha; Glucose > metabolism; Astrocytes > physiology; Brain Injuries > drug therapy
• Online Access: http://library.cuhk.edu.hk/record=b5896406; http://repository.lib.cuhk.edu.hk/en/item/cuhk-325179

Wong Chun Lung. === Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. === Includes bibliographical references (leaves 202-225). === Abstracts in English and Chinese. === Thesis Committee --- p.ii === Abstract --- p.iii === 摘要 --- p.vi === Acknowledgements --- p.ix === Table of Contents --- p.x === List of Abbreviations --- p.xv === List of Figures --- p.xix === List of Tables --- p.xx iii === Chapter Chapter 1 --- Introduction === Chapter 1.1 --- "Neurodegeneration, Inflammation and Gliosis" --- p.1 === Chapter 1.2 --- Anatomy of the CNS --- p.5 === Chapter 1.3 --- Astrocytes --- p.6 === Chapter 1.3.1 --- Morphology and Identification of Astrocytes --- p.6 === Chapter 1.3.2 --- Physiological Functions of Astrocytes in the CNS --- p.7 === Chapter 1.3.2.1 --- Induction of Blood-brain Barrier (BBB) --- p.7 === Chapter 1.3.2.2 --- Metabolism of Neurotransmitters --- p.9 === Chapter 1.3.2.3 --- Nursing Role of Astrocytes --- p.9 === Chapter 1.3.2.4 --- Immunological Functions of Astrocytes --- p.10 === Chapter 1.3.3 --- Neonatal Rat Cortical Astrocytes as In Vitro Model --- p.12 === Chapter 1.4 --- Cytokines in Brain Damage --- p.14 === Chapter 1.4.1 --- Lipopolysaccharides (LPS) --- p.16 === Chapter 1.4.2 --- Tumor Necrosis Factor-α (TNF-α) --- p.17 === Chapter 1.4.3 --- Interleukin-1 (IL-1) --- p.19 === Chapter 1.4.4 --- Interleukin-6 (IL-6) --- p.20 === Chapter 1.4.5 --- Interferon-γ (IFN-γ) --- p.21 === Chapter 1.5 --- Cytokines-induced Signaling Cascade --- p.22 === Chapter 1.5.1 --- TNF Receptors --- p.23 === Chapter 1.5.2 --- Ca2+ --- p.25 === Chapter 1.5.3 --- MAPK --- p.26 === Chapter 1.5.4 --- PICA --- p.27 === Chapter 1.5.5 --- NFkB --- p.29 === Chapter 1.6 --- Glucose Metabolism in the Brain and Glucose Transporters --- p.31 === Chapter 1.6.1 --- Glucose Transporters in the Brain --- p.32 === Chapter 1.6.2 --- Glucose Transporters in Brain Damage --- p.34 === Chapter 1.7 --- Ascorbic Acid Metabolism in the Brain --- p.36 === Chapter 1.8 --- Aim and Scope of this Project --- p.39 === Chapter Chapter 2 --- Materials and Methods === Chapter 2.1 --- Materials === Chapter 2.1.1 --- Neonatal Sprawley 一Dawley Rats --- p.43 === Chapter 2.1.2 --- Plain Dulbecco Modified Eagle Medium ´ؤ Formula 12 (pDF12) --- p.43 === Chapter 2.1.3 --- Complete DF-12(cDF12) --- p.43 === Chapter 2.1.4 --- Phosphate Buffered Saline (PBS) --- p.44 === Chapter 2.1.5 --- Hank's Buffer (HSB) --- p.44 === Chapter 2.1.6 --- D/L-Homocysteine Buffer --- p.44 === Chapter 2.1.7 --- "LPS, Cytokines and Pentoxifylline" --- p.45 === Chapter 2.1.8 --- Specific TNF Receptor Agonist: TNF antibodies --- p.45 === Chapter 2.1.9 --- Calcium Modulators --- p.45 === Chapter 2.1.10 --- PKA Modulators --- p.46 === Chapter 2.1.11 --- NFkB Inhibitors --- p.47 === Chapter 2.1.12 --- MAPK Inhibitors --- p.47 === Chapter 2.1.13 --- β-Adrenergic Receptor Modulators --- p.47 === Chapter 2.1.14 --- Reagents for RNA and Protein Isolation --- p.48 === Chapter 2.1.15 --- Reagents for Reverse Transcription-Polymerase Chain Reaction (RT-PCR) --- p.48 === Chapter 2.1.16 --- Reagents for DNA Electrophoresis --- p.49 === Chapter 2.1.17 --- Reagents for Real-time PCR --- p.51 === Chapter 2.1.18 --- Reagents for Western Blotting --- p.51 === Chapter 2.1.19 --- Reagents for MTT Assay --- p.51 === Chapter 2.1.20 --- Reagents for 3H-Thymidine Incorporation Assay --- p.52 === Chapter 2.1.21 --- Reagents for Glucose Uptake Assay --- p.52 === Chapter 2.1.22 --- Reagents for Ascorbic Acid Accumulation Assay --- p.53 === Chapter 2.1.23 --- Reagents for Immunostammg --- p.53 === Chapter 2.1.24 --- Other Chemicals and Reagents --- p.53 === Chapter 2.2 --- Methods === Chapter 2.2.1 --- Preparation of Primary Cultured Rat Astrocytes --- p.55 === Chapter 2.2.2 --- Measuring Cell Viability: MTT Assay --- p.56 === Chapter 2.2.3 --- Measuring Cell Proliferation: 3H Thymidine Incorporation Assay --- p.57 === Chapter 2.2.4 --- Measuring Glucose Uptake: Zero-trans Glucose Uptake Assay --- p.58 === Chapter 2.2.5 --- Measuring Ascorbic Acid Accumulation --- p.60 === Chapter 2.2.6 --- Total Protein Extraction --- p.61 === Chapter 2.2.7 --- Western Blotting --- p.62 === Chapter 2.2.8 --- Immunostaining --- p.64 === Chapter 2.2.9 --- Isolation of RNA --- p.64 === Chapter 2.2.10 --- Measurement of RNA Yield --- p.65 === Chapter 2.2.11 --- RNA Gel Electrophoresis --- p.66 === Chapter 2.2.12 --- Reverse Transcription (RT) --- p.66 === Chapter 2.2.13 --- Polymerase Chain Reaction (PCR) --- p.67 === Chapter 2.2.14 --- Separation of PCR Products by Agarose Gel Electrophoresis --- p.67 === Chapter 2.2.15 --- Quantization of PCR Products and Western Blotting --- p.68 === Chapter 2.2.16 --- Real-time PCR --- p.68 === Chapter Chapter 3 --- Results === Chapter 3.1 --- Role of Calcium Ions (Ca2+) in TNF-α-induced Astrocyte Proliferation --- p.70 === Chapter 3.1.1 --- Effects of Changes of Extracellular Ca2+ on Astrocyte Viability --- p.72 === Chapter 3.1.2 --- Effects of Other Divalent Ions on Astrocyte Viability --- p.74 === Chapter 3.1.3 --- Effects of Changes of Intracellular Ca2+ on Astrocyte Viability --- p.78 === Chapter 3.1.4 --- Role of Ca2+ on TNF-α-mduced Proliferation in Astrocytes --- p.85 === Chapter 3.1.5 --- Role of Other Divalent Ions on tnf-α-mduced Proliferation in Astrocytes --- p.90 === Chapter 3.2 --- Effect of Cytokines on Glucose Uptake in Rat Astrocytes --- p.95 === Chapter 3.2.1 --- Basal level of Glucose Uptake in Astrocytes and Effects of Cytokines on Glucose Uptake in Astrocytes --- p.95 === Chapter 3.2.2 --- Signaling Cascade of LPS- and TNF-α-induced Glucose Uptake in Astrocytes --- p.120 === Chapter (A) --- TNFR Subtypes Mediating TNF-a-induced Glucose Uptake --- p.121 === Chapter (B) --- MAPK --- p.125 === Chapter (C) --- PKA --- p.133 === Chapter (D) --- NFkB --- p.139 === Chapter (E) --- Other Mechanisms / Signalling molecules --- p.150 === Chapter (1) --- Interaction with β-Adrenegic Mechanism === Chapter (2) --- Role of cGMP --- p.154 === Chapter (3) --- Effect of Mg2+ on LPS- / TNF-α- induced Glucose Uptake in Astrocytes --- p.156 === Chapter (4) --- Possible Involvement of IGF-1 System --- p.160 === Chapter 3.2.3 --- Summary --- p.163 === Chapter 3.3 --- Effects of LPS and Cytokines on AA Accumulation in Astrocytes --- p.164 === Chapter Chapter 4 --- Discussion === Chapter 4.1 --- Role of Calcium ions (Ca2+) in TNF-α-induced Astrocyte Proliferation --- p.177 === Chapter 4.1.1 --- Drastic Changes in Extracellular Ca2+ Caused Astrocyte Death --- p.178 === Chapter 4.1.2 --- Extraordinary Role of Ca2+ in Astrocytes Survival --- p.178 === Chapter 4.1.3 --- Elevation of [Ca2+]i Reduced Astrocyte Viability --- p.180 === Chapter 4.1.4 --- Failure of Verapamil to Block TNF-α-induced Astrocyte Proliferation --- p.182 === Chapter 4.2 --- Hypothesis for the Relationship between Cytokines and Energy Metabolism --- p.185 === Chapter 4.2.1 --- Mechanism and Signaling Cascade of the Elevated Glucose Uptake --- p.186 === Chapter 4.2.2 --- Increased Glucose Uptake by Cytokines: Friend or Foe? --- p.191 === Chapter 4.2.3 --- Depletion of AA Pool by LPS --- p.194 === Chapter 4.2.4 --- Possible Bedside Application of the Findings --- p.195 === Chapter 4.3 --- Prospects of This Study and Concluding Remarks --- p.197 === Appendix --- p.201 === References --- p.202