氟化物对中华大蟾蜍蝌蚪发育影响的研究

摘要 3-6
Abstract 6-9
Chapter 1 Review 12-18
    1.1 Sources of fluoride in the Environment 12
    1.2 Effects of fluoride on organisms 12-13
    1.3 Effects of fluoride on liver 13-14
        1.3.1 Disruption of liver by fluoride 13-14
        1.3.2 Effects of fluoride on lipid metabolism 14
        1.3.3 Effects of fluoride on oxidative stress 14
    1.4 Characterization of lipid metabolism and oxidative stress-related genes 14-18
        1.4.1 Characterization of ACC,FAE, ACOX, SCP, CPT, PPARa 14-16
        1.4.2 Characterization of SOD, GPx and HSPs 16-18
Chapter 2 Effects of fluoride on oxidative stress and lipid metabolismin liver of Bufo gargarizans at pro-metamorphosis stage 18-34
    2.1 Introduction 18-20
    2.2 Materials and methods 20-24
        2.2.1 Animal husbandry 20
        2.2.2 Reagents and solution preparation 20-21
        2.2.3 Experimental design 21
        2.2.4 Histopathological studies 21-22
        2.2.5 Qualitative analysis of melanomacrophage centres and lipid droplets 22
        2.2.6 RNA isolation and cDNA synthesis 22
        2.2.7 Real-time quantitative polymerase chain reaction (RT-qPCR) 22-23
        2.2.8 Statistical analysis 23-24
    2.3 Results 24-30
        2.3.1 Fluoride concentration 24
        2.3.2 Microstructures of the livers 24-26
        2.3.3 Molecular biomarker responses of lipid metabolism 26-27
        2.3.4 Molecular biomarker responses of oxidative stress 27-28
        2.3.5 Molecular biomarker responses of apoptosis 28-30
    2.4 Discussion 30-34
Chapter 3 Effects of fluoride on lipid metabolism,fatty acidscomposition in liver of Bufo gargarizans at metamorphosis climaxstage 34-58
    3.1 Introduction 34-36
    3.2 Materials and methods 36-40
        3.2.1 Test animals 36
        3.2.2 Test solutions 36
        3.2.3 Experimental design 36-37
        3.2.4 Histological analyses of liver 37
        3.2.5 RNA isolation and cDNA synthesis 37
        3.2.6 Real-time quantitative polymerase chain reaction (RT-qPCR) 37-38
        3.2.7 In situ hybridization of liver 38
        3.2.8 Fatty acid extraction and GC-MS analysis 38
        3.2.9 Data analysis 38-40
    3.3 Results 40-52
        3.3.1 Fluoride concentration 40
        3.3.2 Effects of fluoride on the microstructure of liver in B. gargarizans 40-43
        3.3.3 Expression of genes involved in oxidative stress in liver 43-44
        3.3.4 Expression of genes involved in synthesis and secretion of bile acid in liver 44-45
        3.3.5 Expression of genes involved in lipid metabolism in liver 45-48
        3.3.6 In situ hybridization of BSEP 48-49
        3.3.7 In situ hybridization of HSD3B7 49-50
        3.3.8 The effect of fluoride on the composition and content of fatty acids 50-52
    3.4 Discussion 52-58
Chapter 4 Conclusion 58-60
References 60-78
致谢 78-80
攻读硕士学位期间研究成果 80