by Tsang Chi Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 137-155). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abbreviations --- p.iii / Abstract --- p.vi / 撮要 --- p.ix / Publications --- p.xi / Table of contents --- p.xii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Human mast cells and their pathological roles in inflammation --- p.1 / Chapter 1.1.1 --- Morphology of mast cells --- p.1 / Chapter 1.1.2 --- Mediators of mast cells --- p.1 / Chapter 1.1.3 --- Migration and activation --- p.3 / Chapter 1.1.4 --- Pathological roles of mast cells --- p.3 / Chapter 1.1.5 --- Human mast cell-1 (HMC-1) --- p.5 / Chapter 1.2 --- Cytokines as stimulator of mast cells in inflammation --- p.7 / Chapter 1.2.1 --- SCF --- p.7 / Chapter 1.2.2 --- TNF-α --- p.8 / Chapter 1.2.3 --- IL-13 --- p.8 / Chapter 1.2.4 --- IL-18 --- p.9 / Chapter 1.2.5 --- IL-25 --- p.9 / Chapter 1.3 --- Interaction of mast cells with inflammatory cells through adhesion molecules and chemokines --- p.11 / Chapter 1.3.1 --- Adhesion molecules on mast cells --- p.11 / Chapter 1.3.2 --- Chemokines released by mast cells --- p.12 / Chapter 1.4 --- Intracellular signaling pathways in mast cells --- p.16 / Chapter 1.4.1 --- p38-MAPK pathway --- p.16 / Chapter 1.4.2 --- ERK pathway --- p.17 / Chapter 1.4.3 --- NF-kB Pathway --- p.18 / Chapter 1.4.3 --- Cross-talking of pathways --- p.18 / Chapter 1.5 --- Signal transduction pathways and pharmacological intervention --- p.23 / Chapter 1.6 --- Traditional Chinese Medicine and pharmacological intervention --- p.25 / Chapter 1.6.1 --- Anti-allergic effects of traditional Chinese Medicine --- p.25 / Chapter 1.6.2 --- Anti-asthmatic effects of a newly developed Wheeze-Relief Formula --- p.26 / Chapter 1.7 --- Aims and scope of the study --- p.30 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials --- p.32 / Chapter 2.1.1 --- HMC-1 cell Line --- p.32 / Chapter 2.1.2 --- Media and reagents for cell culture --- p.32 / Chapter 2.1.3 --- Recombinant human cytokines --- p.33 / Chapter 2.1.4 --- "Signal transduction pathway inhibitors: PD98035, SB203580 and BAY 117082" --- p.34 / Chapter 2.1.5 --- Monoclonal antibodies and reagents for immunofluorescent staining --- p.34 / Chapter 2.1.6 --- Reagents and buffers for chemokine detection --- p.35 / Chapter 2.1.7 --- Reagents and buffers for total RNA extraction --- p.36 / Chapter 2.1.8 --- Reagents and buffers for reverse transcription 一 polymerase chain reaction (RT-PCR) --- p.37 / Chapter 2.1.9 --- Reagents and buffers for protein extraction --- p.40 / Chapter 2.1.10 --- Reagents and buffers for detection of activated signaling pathways --- p.41 / Chapter 2.1.11 --- Reagents and buffers for agarose gel electrophoresis --- p.42 / Chapter 2.1.12 --- Reagents and buffers for SDS-polyacrylamide gel electrophoresis (PAGE) --- p.43 / Chapter 2.1.13 --- Reagents and buffers for Western blot analysis --- p.45 / Chapter 2.1.14 --- Reagents and buffers for cDNA expression array analysis --- p.47 / Chapter 2.1.15 --- Reagents and buffers for cell viability and proliferation assay --- p.48 / Chapter 2.1.16 --- Reagent kit for endotoxin level assay --- p.49 / Chapter 2.2 --- Methods --- p.49 / Chapter 2.2.1 --- HMC-1 cell cultures --- p.49 / Chapter 2.2.2 --- Flow cytometry of cell surface expression of ICAM-1 and ICAM-3 --- p.50 / Chapter 2.2.3 --- Total cellular RNA extraction --- p.50 / Chapter 2.2.4 --- Reverse Transcription - Polymerase Chain Reaction (RT-PCR) --- p.51 / Chapter 2.2.5 --- Agarose gel electrophoresis --- p.51 / Chapter 2.2.6 --- "Quantitative analysis of IL-8, IP-10,MCP-1 and RANTES" --- p.52 / Chapter 2.2.7 --- Quantitative analysis of 1-309 and MIP-1β --- p.52 / Chapter 2.2.8 --- Detection of phosphorylated-ERX and phosphorylated-p38 MAPK --- p.53 / Chapter 2.2.9 --- Detection of NF-kB activity --- p.53 / Chapter 2.2.10 --- Detection of phosphorylated-ATF-2 --- p.53 / Chapter 2.2.11 --- Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) --- p.54 / Chapter 2.2.12 --- Western blot analysis --- p.54 / Chapter 2.2.13 --- MTT assay --- p.55 / Chapter 2.2.14 --- Cell proliferation assay --- p.55 / Chapter 2.2.15 --- Hot water extraction of TCM --- p.56 / Chapter 2.2.16 --- Endotoxin level assay --- p.56 / Chapter 2.2.17 --- cDNA expression array analysis --- p.57 / Chapter 2.2.18 --- Statistical analysis --- p.57 / Chapter Chapter 3 --- Results / Chapter 3.1 --- The effects of cytokines on the expression of ICAM-1 and ICAM-3 on HMC-1 --- p.59 / Chapter 3.1.1. --- "SCF, TNF-α and IL-13 up-regulated ICAM-1 but not ICAM-3 expression on HMC-1 cells" --- p.59 / Chapter 3.1.2. --- "SCF, TNF-α and IL-13 up-regulated the mRNA expression of ICAM-1" --- p.59 / Chapter 3.1.3 --- "The combined treatment of SCF and TNF-α, and SCF and IL-13 showed synergistic and additive effect on ICAM-1 expression respectively" --- p.60 / Chapter 3.1.4 --- Synergistic up-regulation of ICAM-1 expression in combined treatment of SCF and TNF-α was dose-dependently enhanced by SCF --- p.60 / Chapter 3.2 --- "The effects of cytokines on the release of IL-8, IP-10, MCP-1, RANTES, 1-309 and MIP-1β from HMC-1 cells" --- p.66 / Chapter 3.2.1 --- "SCF induced the release of IL-8, MCP-1, RANTES, 1-309 and MIP-1β" --- p.66 / Chapter 3.2.2 --- "TNF-a induced the release of IL-8, IP-10, MCP-1, RANTES and 1-309" --- p.66 / Chapter 3.2.3 --- SCF and TNF-α did not enhance the proliferation rate of HMC-1 --- p.66 / Chapter 3.3 --- "The effect of SCF and TNF-α on the activation of ERK, p38 MAPK and NK-kB" --- p.71 / Chapter 3.3.1 --- SCF activated ERK but not p38 MAPK and NF-kB --- p.71 / Chapter 3.3.2 --- TNF-α activated p38 MAPK and NF-kB but not ERK --- p.71 / Chapter 3.4 --- The effect of inhibitors on the SCF and TNF-a-induced release of chemokines --- p.76 / Chapter 3.4.1 --- "The optimal dose of PD98059, SB203580 and BAY117082" --- p.76 / Chapter 3.4.2 --- "PD98059 suppressed the SCF induced IL-8, MCP-1, RANTES, 1-309 and MIP-1β release from HMC-1 cells" --- p.76 / Chapter 3.4.3 --- SB203580 and BAY117082 differentially suppressed the TNF-α induced chemokine release from HMC-1 cells --- p.77 / Chapter 3.5 --- The effect of inhibitors on the SCF and TNF-a-induced upregulation of ICAM-1 --- p.83 / Chapter 3.5.1 --- BAY117082 but not SB203580 suppressed the TNF-α-induced ICAM-1 expression --- p.83 / Chapter 3.5.2 --- PD98059 and BAY 117082 suppressed the combined treatment of SCF and TNF-α induced ICAM-1 expression --- p.83 / Chapter 3.6 --- "Effect of inhibitors on TNF-α and SCF-induced ERK, p38 MAPK and NF-kB activities in HMC-1 cells." --- p.85 / Chapter 3.6.1 --- PD98059 suppressed the SCF-induced activity of ERK --- p.85 / Chapter 3.6.2 --- SB203580 and BAY117082 suppressed the TNF-α induced p38 MAPKand NF-kB activity respectively --- p.85 / Chapter 3.6.3 --- PD98059 suppressed the enhanced NF-kB activity after the combined treatment of SCF and TNF-α for 18 hours --- p.86 / Chapter 3.7 --- Effect of TNF-α and SCF on the gene expression profile of inflammatory cytokines and receptors of HMC-1 cells. --- p.90 / Chapter 3.8 --- The effects of TCM on the SCF-induced 1-309 and MCP-1 from HMC-1 cells --- p.95 / Chapter 3.8.1 --- "Endotoxin level of Radix astragali, Radix Scutellariae, Radix stemonae, Bulbus Fritillariae cirrhosae and Cordyceps sinensis" --- p.95 / Chapter 3.8.2 --- The effects of TCM on the proliferation rate of HMC-1 cells --- p.95 / Chapter 3.9.3 --- The effects of TCM on the SCF-induced release of 1-309 from HMC-1 cells --- p.96 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- Involvement of adhesion molecules and chemokines in mast cell-mediated immunological events --- p.107 / Chapter 4.2 --- HMC-1 as the in vitro mast cell model adapted in my project --- p.108 / Chapter 4.3 --- The effect of cytokines on the expression of ICAM-1 and ICAM-3 in HMC-1 cells --- p.109 / Chapter 4.4 --- The effect of cytokines on the release of chemokines in HMC-1 cells --- p.111 / Chapter 4.5 --- "The regulation of ICAM-1, IL-8, IP-10, MCP-1, RANTES, 1-309 and MIP-1β through p-38 MAPK, ERK and NF-kB signaling pathways in HMC-1 cells" --- p.115 / Chapter 4.6 --- Further characterization of HMC-1 cells using cDNA array --- p.119 / Chapter 4.7 --- Investigating the in vitro anti-allergic activities of a newly developed Wheeze-relief formula using cytokine-activated HMC-1 cells --- p.128 / Chapter 4.8 --- Concluding remarks and future prospective --- p.132 / References --- p.137 / Appendix --- p.156
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_325265 |
| Date | January 2005 |
| Contributors | Tsang, Chi Man., Chinese University of Hong Kong Graduate School. Division of Chinese Medicine. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xvi, 159 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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