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

• Other Authors: Guo, Cuixia.
• Format: Others
• Language: English; Chinese
• Published: 2010
• Subjects: Edible mushrooms; Sclerotium (Mycelium); Immune response > Regulation; Macrophages
• Online Access: http://library.cuhk.edu.hk/record=b5894347; http://repository.lib.cuhk.edu.hk/en/item/cuhk-327243

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