Investigations on the anti-diabetic activities of traditional Chinese medicine formulae originally used against diabetic foot ulcer.

Chan Chak Ming.Thesis (M.Phil.)--Chinese University of Hong Kong, 2004.Includes bibliographical references (leaves 178-202).Abstracts in English and Chinese.Abstract --- p.iAbstract in Chinese 摘耍 --- p.iiiAcknowledgements --- p.vTable of contents --- p.viList of tables --- p.xiiList of figures --- p.xiiiAbbreviations --- p.xviChapter Chapter 1: --- Introduction --- p.1Chapter 1.1 --- Definition of diabetes mellitus --- p.1Chapter 1.2 --- Classification of diabetes mellitus --- p.4Chapter 1.2.1 --- Type 1 diabetes --- p.4Chapter 1.2.2 --- Type 2 diabetes --- p.5Chapter 1.2.3 --- Other forms of diabetes --- p.9Chapter 1.3 --- Complications of diabetes mellitus --- p.11Chapter 1.4 --- Current treatment of diabetes mellitus --- p.12Chapter 1.4.1 --- Type 1 diabetes --- p.12Chapter 1.4.2 --- Type 2 diabetes --- p.13Chapter 1.4.2.1 --- Diet and exercise --- p.13Chapter 1.4.2.2 --- Medication --- p.13Chapter 1.5 --- The use of herbal medicines in diabetes treatment --- p.18Chapter 1.6 --- "Hypothesis, objectives and design of the project" --- p.22Chapter Chapter 2: --- Preparation and authentication of traditional Chinese medicines --- p.23Chapter 2.1 --- Introduction --- p.23Chapter 2.1.1 --- Background information of the formulae --- p.23Chapter 2.1.2 --- Component herbs of formula1 --- p.26Chapter 2.2 --- Objectives --- p.29Chapter 2.3 --- Materials --- p.30Chapter 2.3.1 --- Raw herbal materials and formula 1 extract --- p.30Chapter 2.3.2 --- Thin layer chromatography --- p.35Chapter 2.3.3 --- High performance liquid chromatography determination of the sugar content of the herbal extracts --- p.38Chapter 2.4 --- Methods --- p.41Chapter 2.4.1 --- Thin layer chromatography of the component herbs --- p.41Chapter 2.4.2 --- Raw herbal materials water extraction --- p.46Chapter 2.4.3 --- High performance liquid chromatography determination of the sugar content of the herbal extracts --- p.46Chapter 2.5 --- Results --- p.49Chapter 2.5.1 --- Thin layer chromatography of the component herbs --- p.49Chapter 2.5.2 --- Raw herbal materials water extraction --- p.56Chapter 2.5.3 --- High performance liquid chromatography determination of the sugar content of the herbal extracts --- p.57Chapter 2.6 --- Discussion --- p.62Chapter Chapter 3: --- The anti-diabetic effects of formula 1 and its component herbs in vitro --- p.67Chapter 3.1 --- Introduction --- p.67Chapter 3.1.1 --- Glycaemic control in type 2 diabetes --- p.67Chapter 3.1.2 --- Type 2 diabetes and peripheral tissues --- p.70Chapter 3.1.3 --- Type 2 diabetes and liver --- p.73Chapter 3.1.4 --- Type 2 diabetes and intestinal glucose absorption --- p.76Chapter 3.2 --- Objectives --- p.79Chapter 3.3 --- Materials --- p.80Chapter 3.3.1 --- Cell lines --- p.80Chapter 3.3.2 --- "Cell culture media, buffers, reagents and culture wares" --- p.81Chapter 3.3.3 --- "Chemicals, media and reagents for 3T3-L1 differentiation" --- p.83Chapter 3.3.4 --- Chemicals and reagents for 3T3-L1 and Hs68 2-deoxy-D- glucose (2-DG) uptake assay --- p.84Chapter 3.3.5 --- Chemicals and buffers for H4IIE glucose production assay and phosphoenolpyruvate carboxykinase (PEPCK) assay --- p.85Chapter 3.3.6 --- "Animal, buffers and reagents for preparation and glucose uptake assay of brush border membrane vesicles (BBMV)" --- p.87Chapter 3.3.7 --- Reagents for bicinchoninic acid (BCA) protein assay --- p.88Chapter 3.4 --- Methods --- p.89Chapter 3.4.1 --- Cell culture --- p.89Chapter 3.4.2 --- Studies on glucose uptake in 3T3-L1 adipocytes and Hs68 fibroblasts --- p.90Chapter 3.4.2.1 --- Differentiation of 3T3-L1 cells --- p.90Chapter 3.4.2.2 --- Oil red O staining of the 3T3-L1 cells --- p.90Chapter 3.4.2.3 --- 2-DG uptake assay of 3T3-L1 adipocytes and Hs68 fibroblasts --- p.91Chapter 3.4.3 --- Studies on gluconeogenesis in H4IIE hepatoma cells --- p.93Chapter 3.4.3.1 --- Glucose production assay --- p.93Chapter 3.4.3.2 --- PEPCK assay --- p.94Chapter 3.4.4 --- Studies on BBMV glucose uptake --- p.95Chapter 3.4.4.1 --- Preparation of BBMV --- p.95Chapter 3.4.4.2 --- Preparation of the chloroform extract of the herbal water extract --- p.96Chapter 3.4.4.3 --- Glucose uptake assay of BBMV --- p.97Chapter 3.4.5 --- BCA (Bicinchoninic acid) protein assay --- p.99Chapter 3.4.6 --- Statistical analysis --- p.100Chapter 3.5 --- Results --- p.101Chapter 3.5.1 --- Glucose uptake assay in 3T3-L1 adipocytes and Hs68 fibroblasts --- p.101Chapter 3.5.2 --- Glucose production and PEPCK assay in H4IIE hepatoma cells --- p.108Chapter 3.5.3 --- Glucose uptake assay in BBMV --- p.113Chapter 3.6 --- Discussion --- p.119Chapter 3.6.1 --- Glucose uptake in 3T3-L1 adipocytes and Hs68 fibroblasts --- p.119Chapter 3.6.2 --- Glucose production and PEPCK activity in H4IIE hepatoma cells --- p.123Chapter 3.6.3 --- Glucose absorption in BBMV --- p.125Chapter 3.6.4 --- Conclusion --- p.128Chapter Chapter 4: --- The anti-diabetic effects of formula 1 and Rhizoma Smilacis Chinensis in vivo --- p.131Chapter 4.1 --- Introduction --- p.131Chapter 4.1.1 --- Diabetic animal models --- p.131Chapter 4.1.2 --- Neonatal streptozotocin-induced diabetic rat model --- p.133Chapter 4.2 --- Objective --- p.136Chapter 4.3 --- Materials --- p.137Chapter 4.3.1 --- Animals --- p.137Chapter 4.3.2 --- Chemicals and reagent kit --- p.137Chapter 4.4 --- Methods --- p.139Chapter 4.4.1 --- Induction of diabetes in rats --- p.139Chapter 4.4.2 --- Oral glucose tolerance test --- p.139Chapter 4.4.3 --- Basal glycaemia test --- p.141Chapter 4.4.4 --- Plasma glucose level determination --- p.142Chapter 4.4.5 --- Statistical analysis --- p.142Chapter 4.5 --- Results --- p.143Chapter 4.5.1 --- Oral glucose tolerance test --- p.143Chapter 4.5.2 --- Basal glycaemia test --- p.147Chapter 4.6 --- Discussion --- p.151Chapter Chapter 5: --- The effects of the TCM treatment on glucose homeostasis in diabetic foot ulcer patients --- p.155Chapter 5.1 --- Introduction --- p.155Chapter 5.2 --- Objective --- p.156Chapter 5.3 --- Materials --- p.157Chapter 5.3.1 --- Study subjects --- p.157Chapter 5.3.2 --- Blood sample --- p.158Chapter 5.3.3 --- Chemicals and reagents for erythrocyte glucose uptake assay --- p.158Chapter 5.4 --- Methods --- p.160Chapter 5.4.1 --- Preparation of blood sample --- p.160Chapter 5.4.2 --- Zero-trans influx of 3-OMG uptake in erythrocytes --- p.160Chapter 5.4.3 --- Statistical analysis --- p.161Chapter 5.5 --- Results --- p.162Chapter 5.6 --- Discussion --- p.166Chapter Chapter 6: --- General discussion and conclusion --- p.168Chapter 6.1 --- Overview of the project and analysis of research findings --- p.168Chapter 6.2 --- Limitations of the study --- p.173Chapter 6.3 --- Future directions --- p.174Chapter 6.4 --- Conclusion --- p.177Chapter Chapter 7: --- References --- p.177Appendices --- p.203Appendix I The determination of the sugar contents in the herbal water extracts by high performance liquid chromatography --- p.203Appendix II Basal glycaemia test of formula 1 (822mg/kg) on nO-STZ rats --- p.20