Immunopathological mechanisms of inflammatory reaction in Chinese patients with type 2 diabetic nephropathy: clinical and in vitro studies.

Abstract

Ho, Wing-Yin.Thesis (M.Phil.)--Chinese University of Hong Kong, 2007.Includes bibliographical references (leaves 115-131).Abstracts in English and Chinese.Acknowledgements --- p.iAbbreviations --- p.iiiAbstract --- p.v摘要 --- p.ixPublications --- p.xiiTable of Contents --- p.xivChapter 1. --- General IntroductionChapter 1.1. --- Diabetes Mellitus (DM) and Diabetic Nephropathy --- p.1Chapter 1.1.1. --- "Prevalence, Diagnosis and Classification of DM" --- p.1Chapter 1.1.2. --- Type 2 DM and its Complications: Diabetic Nephropathy --- p.5Chapter 1.1.3. --- Diagnosis and Impacts of Diabetic Nephropathy --- p.7Chapter 1.1.4. --- Current Treatment of Type 2 DM and Diabetic Nephropathy --- p.8Chapter 1.2. --- Cytokines and Chemokines --- p.9Chapter 1.2.1. --- Types and Properties --- p.9Chapter 1.2.2. --- Cytokines and chemokines in Type 2 DM and Diabetic Nephropathy --- p.13Chapter 1.3. --- T Lymphocyte Costimulatory Molecules --- p.15Chapter 1.3.1. --- Types and Properties --- p.15Chapter 1.3.2. --- T Lymphocyte Costimulatory Molecules in Type 2 DM and Diabetic Nephropathy --- p.16Chapter 1.4. --- Adhesion Molecules --- p.18Chapter 1.4.1. --- Types and Properties --- p.18Chapter 1.4.2. --- Adhesion Molecules in Type 2 DM and Diabetic Nephropathy --- p.20Chapter 1.5. --- Intracellular Signaling Pathways --- p.21Chapter 1.5.1. --- Types and Properties --- p.21Chapter 1.5.2. --- Intracellular Signaling Pathways in Type 2 DM and Diabetic Nephropathy --- p.23Chapter 1.6. --- Objectives of Our Study --- p.24Chapter 2. --- Materials and MethodsChapter 2.1. --- Materials --- p.26Chapter 2.1.1. --- "Patients, Control Subjects and Blood Samples" --- p.26Chapter 2.1.2. --- Cell Line --- p.27Chapter 2.1.3. --- "Cell Culture Media, Buffers and Other Reagents" --- p.28Chapter 2.1.4. --- "Recombinant Human Cytokines, Inhibitors and Other Stimulators" --- p.30Chapter 2.1.5. --- Reagents and Buffers for Flow Cytometric Analysis --- p.31Chapter 2.1.5.1. --- Cytometric Bead Array (CBA) of Cytokines and Chemokines --- p.33Chapter 2.1.5.2. --- Multiplex Fluorescent Bead Immunoassay (FBI) of Soluble Adhesion Molecules --- p.33Chapter 2.1.5.3. --- Phosphorylation State Analysis of Signaling Molecules --- p.34Chapter 2.1.5.4. --- Immunofluorescent Staining of Cell Surface Molecules --- p.36Chapter 2.1.6. --- Reagents and Buffers for Protein Array Analysis --- p.37Chapter 2.1.7. --- "Reagents and Buffers for 3-(4,5-Dimethylthiazol-2-yl)-2,5- diphenylytetrazolium Bromide (MTT) Assay" --- p.37Chapter 2.1.8. --- Reagents for Human Enzyme-Linked Immunosorbent Assay (ELISA) --- p.37Chapter 2.2. --- Methods --- p.38Chapter 2.2.1. --- Whole Blood Culture Experiments --- p.38Chapter 2.2.2. --- "Collection of Serum and Plasma, and Purification of PBMC from EDTA-Blood" --- p.39Chapter 2.2.3. --- HK-2 Cell Cultures --- p.39Chapter 2.2.4. --- HK-2 Cell Treatments --- p.40Chapter 2.2.5. --- Flow Cytometric Analysis --- p.41Chapter 2.2.5.1. --- CBA of Cytokines and Chemokines --- p.41Chapter 2.2.5.2. --- Multiplex FBI of Soluble Adhesion Molecules --- p.41Chapter 2.2.5.3. --- Phosphorylation State Analysis of Signaling Molecules --- p.42Chapter 2.2.5.4. --- Immunofluorescent Staining of Cell Surface Molecules --- p.43Chapter 2.2.6. --- Protein Array Analysis --- p.44Chapter 2.2.7. --- MTT Assay --- p.44Chapter 2.2.8. --- ELISA --- p.45Chapter 2.2.9. --- Statistical Analysis --- p.46Chapter 3. --- "Clinical Study on the Expressions of Cytokines, Chemokines, Co-stimulatory Molecules, Phosphorylated Signaling Molecules in Patients with Diabetic Nephropathy"Chapter 3.1. --- Introduction --- p.47Chapter 3.2. --- Results --- p.49Chapter 3.2.1. --- Demographic Data of Participants --- p.49Chapter 3.2.2. --- Expression Profile in Plasma of Patients --- p.49Chapter 3.2.2.1. --- Cytokines and Chemokines --- p.49Chapter 3.2.2.2. --- Soluble Costimulatory Molecules --- p.55Chapter 3.2.2.3. --- Soluble Adhesion Molecules --- p.55Chapter 3.2.2.4. --- "Correlations between Plasma Concentrations of Cytokines, Chemokines, soluble Costimulatory Molecules and soluble Adhesion Molecules and UACR in Patients" --- p.60Chapter 3.2.3. --- Effects ofTNF-α and IL-18 on the ex vivo Production from Whole Blood of Patients --- p.65Chapter 3.2.3.1. --- Ex vivo Production of Cytokines and Chemokines --- p.65Chapter 3.2.3.2. --- Ex vivo Production of Soluble Costimulatory Molecules --- p.70Chapter 3.2.4. --- "Expression of Phosphorylated p38 MAPK, JNK and ERK in PBMC of Patients" --- p.73Chapter 3.3. --- Discussion --- p.77Chapter 3.3.1. --- "Cytokines, Chemokines and Diabetic Nephropathy" --- p.77Chapter 3.3.2. --- Soluble Costimulatory Molecules and Diabetic Nephropathy --- p.80Chapter 3.3.3. --- Soluble Adhesion Molecules and Diabetic Nephropathy --- p.83Chapter 3.3.4. --- Intracellular Signaling and Diabetic Nephropathy --- p.87Chapter 4. --- In vitro Study on the Signal Transduction Mechanism Regulating the Expression of CCL2 and Cell Surface Adhesion Molecules in Tumour Necrosis Factor (TNF)-α-Stimulated HK-2 CellsChapter 4.1. --- Introduction --- p.90Chapter 4.2. --- Results --- p.93Chapter 4.2.1. --- Expression Profile of Cytokines and Chemokines of TNF-α-activated HK-2 Cells --- p.93Chapter 4.2.2. --- "TNF-α Upregulated CCL2, ICAM-1 and VCAM-1 Expression in HK-2 Cells" --- p.95Chapter 4.2.3. --- "TNF-α Activated the p38 MAPK, JNK and ERK Signaling Pathways in HK-2 Cells" --- p.96Chapter 4.2.4. --- Cytotoxicity of MAPK Inhibitors --- p.96Chapter 4.2.5. --- "Effects of p38 MAPK, JNK and ERK Inhibitors on TNF-α-induced Expressions of CCL2, ICAM-1 and VCAM-1" --- p.100Chapter 4.3. --- Discussion --- p.102Chapter 5. --- Conclusion and Future ProspectsChapter 5.1. --- Conclusion --- p.107Chapter 5.2. --- Future Prospects --- p.111References --- p.11