Antioxidant and immunomodulatory properties from plant materials (studies on ganoderma lucidum).

by Fu Sai-chuen.Thesis (M.Phil.)--Chinese University of Hong Kong, 1994.Includes bibliographical references (leaves 102-111).AcknowledgementsList of AbbreviationAbstractPrologue --- p.1ChapterChapter 1. --- Introduction --- p.2Chapter 1.1 --- Historical background of Ling Zhi (靈芝） --- p.4Chapter 1.2 --- Biological description of Ling Zhi (靈芝） --- p.6Chapter 1.3 --- Chemical composition of Ganoderma lucidum --- p.6Chapter 1.3.1 --- The triterpenoids --- p.7Chapter 1.3.2 --- Matrix polysaccharides --- p.8Chapter 1.4 --- Medicinal properties and biological activities --- p.9Chapter 1.4.1 --- Medicinal properties --- p.9Chapter 1.4.2 --- Biological activities --- p.10Chapter 1.4.2.1 --- Anti-tumour activity --- p.10Chapter 1.4.2.2 --- Enhancement of protein and nucleic acid synthesis --- p.11Chapter 1.4.2.3 --- Effects on nervous system --- p.11Chapter 1.4.2.4 --- Effects on respiratory system --- p.12Chapter 1.4.2.5 --- Effects on cardiovascular system --- p.12Chapter 1.4.2.6 --- Effects on immune system --- p.14Chapter 1.4.2.7 --- Hepatoprotection and detoxicant actions --- p.14Chapter 1.4.3 --- Summary --- p.15Chapter 1.5 --- The relationship between antioxidant defense/immune system and disease --- p.15Chapter 2. --- Materials and Methods --- p.20Chapter 2.1 --- Cell culture reagentsChapter 2.1.1 --- Preparation of RPMI-1640 medium --- p.20Chapter 2.1.2 --- Preparation of fetal calf serum (FCS) --- p.20Chapter 2.1.3 --- Preparation of phosphate-buffered saline (PBS) --- p.20Chapter 2.1.4 --- Mitogens and test samples --- p.21Chapter 2.1.5 --- Trypan blue exclusion test --- p.21Chapter 2.1.6 --- Liquid scintillation counting --- p.21Chapter 2.2 --- Animal care --- p.22Chapter 2.3 --- Fractionation of Ganoderma lucidum extracts --- p.22Chapter 2.4 --- Carbon tetrachloride (CC14)- induced hepatotoxicity in mice --- p.23Chapter 2.4.1 --- Pretreatment scheme and CCl4 treatment --- p.23Chapter 2.4.2 --- Preparation of liver homogenates --- p.23Chapter 2.4.3 --- Biochemical assays for assessing hepatotoxicity --- p.25Chapter 2.4.3.1 --- Determination of plasma alanine aminotransferase (ALT) --- p.25Chapter 2.4.3.2 --- Determination of tissue glutathione (GSH) content --- p.25Chapter 2.4.3.3 --- Depletion of tissue GSH in liver homogenates by tert- butylhydroperoxide (tBHP) --- p.26Chapter 2.4.3.4 --- Determination of tissue malondialdehyde (MDA) content --- p.26Chapter 2.5 --- Preparation of murine hepatocytes --- p.27Chapter 2.5.1 --- Liver perfusion --- p.27Chapter 2.5.2 --- Isolation of murine hepatocytes --- p.29Chapter 2.5.3 --- Incubation protocol --- p.29Chapter 2.5.4 --- Determination of cellular glutathione by HPLC method --- p.30Chapter 2.6 --- Preparation of splenocytes and lymphocytes --- p.31Chapter 2.6.1 --- Isolation of murine splenocytes --- p.31Chapter 2.6.2 --- Mitogenic assay by measuring 3H-thymidine uptake --- p.31Chapter 2.6.3 --- B cell- and T cell- enrichment in splenocyte preparation --- p.32Chapter 2.6.4 --- Isolation of human peripheral blood lymphocytes (HPBL) from cord blood --- p.34Chapter 2.7 --- Chromatographic methods --- p.34Chapter 2.7.1 --- Fast Protein Liquid Chromatography (FPLC) with Superose 12 column --- p.34Chapter 2.7.2 --- Removal of lipopolysaccharide (LPS) with Detoxi Gel --- p.35Chapter 2.7.3 --- Desalting by Sephadex G-25 --- p.35Chapter 2.7.4 --- Determination of FDNB-conjugated GSH and GSSG by HPLC reverse phase chromatography --- p.36Chapter 2.8 --- Measurement of in vivo antibody production --- p.39Chapter 2.8.1 --- Immunization and pretreatment scheme --- p.39Chapter 2.8.2 --- Direct plaque assay --- p.39Chapter 2.9 --- Biochemical analysis --- p.39Chapter 2.9.1 --- Determination of protein content --- p.40Chapter 2.9.2 --- Determination of hexose content --- p.40Chapter 2.9.3 --- Determination of uronic acid content --- p.40Chapter 2.9.4 --- Determination of sulphate content --- p.41Chapter 2.9.5 --- Determination of hexosamine content --- p.42Chapter 2.10 --- Statistical analysis --- p.42Chapter 3. --- Results --- p.44Chapter 3.1 --- Extraction and fractionation of Ganoderma lucidum --- p.44Chapter 3.2 --- Hepatoprotective effect of Ganoderma lucidum fractions aganist CCl4-induced hepatotoxicity --- p.47Chapter 3.2.1 --- Hepatoprotection of Ganoderma lucidum fractions --- p.47Chapter 3.2.2 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic malondialdehyde (MDA) level in CCl4-treated mice --- p.47Chapter 3.2.3 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic glutathione (GSH) level in CCl4-treated mice --- p.51Chapter 3.3 --- Effect of GL2 on isolated hepatocytes --- p.51Chapter 3.3.1 --- Protection against phorone-induced hepatocytotoxicity --- p.51Chapter 3.3.2 --- Protection against menadione-induced hepatocytotoxicity --- p.55Chapter 3.4 --- Immunomodulatory effect of Ganoderma lucidum fractions --- p.55Chapter 3.4.1 --- Discovery of a mitogenic principle from Ganoderma lucidum fractions --- p.55Chapter 3.4.2 --- B cell-specific mitogenic activity --- p.58Chapter 3.4.3 --- Mitogenic activity in Ganoderma lucidum distinguishable from possible LPS contamination --- p.65Chapter 3.4.3.1 --- The use of polymyxin B sulphate --- p.65Chapter 3.4.3.2 --- The use of Detoxi Gel --- p.69Chapter 3.4.3.3 --- The combined use of polymyxin B sulphate and Detoxi Gel --- p.69Chapter 3.5 --- Enhancement of humoral response to sheep red blood cells (SRBC) by the mitogenic GL2 fraction --- p.72Chapter 3.6 --- Physical-chemical characterization of GL2 fraction --- p.76Chapter 3.6.1 --- Estimation of molecular size by FPLC size exclusion chromatography with Superose 12 column --- p.76Chapter 3.6.2 --- Sugar composition of GL2 fraction --- p.76Chapter 3.6.3 --- Comparison of elution profiles in Superose12 column with respect to mitogenic activity and sugar composition --- p.80Chapter 4. --- Discussions --- p.83Chapter 4.1 --- Enhancement of antioxidant status by Ganoderma lucidum fractions --- p.83Chapter 4.2 --- The significance of enhancement of GSH status --- p.89Chapter 4.3 --- Immunomodulatory actions of Ganoderma lucidum --- p.95Conclusions --- p.99Appendix --- p.100Reference --- p.10