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Immunomodulatory effects of hot water extracts isolated from mushroom sclerotia on the biological functions of murine macrophages.

Guo, Cuixia. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 75-85). / Abstracts in English and Chinese. / Thesis committee --- p.ii / Abstract --- p.iii / 摘要 --- p.iv / Acknowledgment --- p.v / List of Tables --- p.vi / List of Figures --- p.vii / List of Abbreviations --- p.viii / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Introduction to immune system --- p.1 / Chapter 1.2 --- Immune effecter cells --- p.1 / Chapter 1.2.1 --- Macrophage --- p.1 / Chapter 1.2.2 --- Dendritic Cells (DCs) --- p.5 / Chapter 1.3 --- Immunomodulatory and antitumor activities of mushrooms --- p.8 / Chapter 1.3.1 --- Introduction to mushroom --- p.11 / Chapter 1.3.2 --- Mushroom polysaccharides --- p.11 / Chapter 1.3.3 --- Mushroom β-glucan --- p.14 / Chapter 1.4 --- The receptors for polysaccharides associated with immune effecter cells --- p.16 / Chapter 1.4.1 --- CR3 --- p.16 / Chapter 1.4.2 --- Dectin-1 --- p.18 / Chapter 1.4.3 --- TLR2 --- p.19 / Chapter 1.5 --- Nuclear factor-kappa B (NF-kB) activation --- p.19 / Chapter 1.6 --- Previous studies on mushroom sclerotium --- p.20 / Chapter 1.6.1 --- Pleurotus tuber-regium (PT) --- p.20 / Chapter 1.6.2 --- Polyporus rhinocerus (PR) --- p.21 / Chapter 1.7 --- Objectives --- p.21 / Chapter 2. --- Materials and Methods --- p.23 / Chapter 2.1 --- Materials --- p.23 / Chapter 2.1.1 --- Mushroom sclerotia --- p.23 / Chapter 2.1.2 --- Animal --- p.23 / Chapter 2.1.3 --- Cell lines --- p.24 / Chapter 2.2 --- Methods --- p.24 / Chapter 2.2.1 --- Hot water extraction --- p.24 / Chapter 2.2.2 --- Measurement of monosaccharide profile --- p.25 / Chapter 2.2.2.1 --- Acid depolymerization --- p.25 / Chapter 2.2.2.2 --- Neutral sugar derivatization --- p.25 / Chapter 2.2.2.3 --- Gas chromatography (GC) --- p.26 / Chapter 2.2.3 --- Determination of molecular weight by size exclusion chromatography (SEC) --- p.27 / Chapter 2.2.4 --- Determination of total sugar by phenol-sulfuric acid method --- p.28 / Chapter 2.2.5 --- Determination of protein content by Lowry-Folin method --- p.28 / Chapter 2.2.6 --- Detection of endotoxin --- p.29 / Chapter 2.2.7 --- Immunomodulatory activities induced in RAW264.7 cell line and murine peritoneal macrophages (PMs) --- p.30 / Chapter 2.2.7.1 --- Isolation of murine peritoneal macrophages (PMs) --- p.30 / Chapter 2.2.7.2 --- Detection of cell surface antigens on RAW 264.7 cells and PMs --- p.30 / Chapter 2.2.7.3 --- Phagocytic uptake --- p.31 / Chapter 2.2.7.4 --- Reactive Oxygen Species (ROS) generation --- p.32 / Chapter 2.2.7.5 --- Nitric Oxide (NO) production --- p.32 / Chapter 2.2.7.6 --- Inducible Nitric Oxide Synthase (iNOS) expression --- p.32 / Chapter 2.2.7.6.1 --- Cell lysates preparation --- p.33 / Chapter 2.2.7.6.2 --- Determination of protein concentrations --- p.33 / Chapter 2.2.7.6.3 --- Western blot --- p.34 / Chapter 2.2.7.7 --- Tumor Necrosis Factor-alpha (TNF-α) production --- p.36 / Chapter 2.2.8 --- DC cell marker determination --- p.37 / Chapter 2.2.9 --- Nuclear factor kappa B (NF-kB) activation --- p.37 / Chapter 2.2.10 --- Determination of the expression of existing cell surface β-glucan receptors --- p.37 / Chapter 2.2.11 --- Statistical methods --- p.38 / Chapter 3. --- Results --- p.39 / Chapter 3.1 --- Yield and chemical composition of mushroom sclerotial polysaccharides --- p.39 / Chapter 3.2 --- Endotoxin examination --- p.41 / Chapter 3.3 --- Monosaccharide profiles of PTW and PRW by GC --- p.41 / Chapter 3.4 --- Molecular weight profile by size exclusion chromatography (SEC) --- p.43 / Chapter 3.5 --- Immunomodulatory activities induced in RAW264.7 cells and murine peritoneal macrophages (PMs) --- p.46 / Chapter 3.5.1 --- Detection of cell surface antigens on RAW 264.7 cells and PMs --- p.46 / Chapter 3.5.2 --- Phagocytic uptake --- p.49 / Chapter 3.5.3 --- ROS generation --- p.53 / Chapter 3.5.4 --- NO production --- p.56 / Chapter 3.5.5 --- iNOS expression --- p.59 / Chapter 3.5.6 --- TNF-α production --- p.60 / Chapter 3.5.7 --- Morphological changes of cells --- p.62 / Chapter 3.5.8 --- DC cell marker determination --- p.64 / Chapter 3.6 --- Receptors expression on RAW 264.7 cells and PMs --- p.66 / Chapter 3.7 --- NF-kB activation --- p.68 / Chapter 3.8 --- Discussion --- p.70 / Chapter 4. --- Conclusions and Future Works --- p.73 / Chapter 5. --- References --- p.75

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_327243
Date January 2010
ContributorsGuo, Cuixia., Chinese University of Hong Kong Graduate School. Division of Life Sciences.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xii, 85 leaves : ill. (chiefly col.) ; 30 cm.
RightsUse 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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