臺灣博碩士論文加值系統

CONTENTS
Contents i
Abstract in Chinese viii
Abstract in English x
List of Tables xiii
List of Figures xiv
Abbreviations xvii
CHAPTER 1. INTRODUCTION
1.1 Background Study 2
1.2 Objectives 8
CHAPTER II. LITERATURE REVIEW
2.1 Sweet Potato 10
2.1.1 Classification 11
2.1.2 Agricultural Profile 12
2.1.3 Origin, Distributin, and Diversity 13
2.1.4 Functional Uses 15
2.1.5 Phytochemicals 16
2.1.5.1 Phenolics 16
2.1.5.2 Flavonoids 19
2.1.5.3 Anthocyanins 21
2.2 Purple-Fleshed Sweet Potato 31
2.3 Purple-Fleshed Sweet Potato (PFSP) Trends 32
2.4 Free Radicals 35
2.4.1 Biological Roles of Free Radicals 35
2.4.1.1 Nitric Oxide as a Free Radical 36
2.4.1.2 Superoxide and Hydrogen Peroxide Radicals 38
2.4.1.3 Iron as Free Radicals 40
2.5 Skin Tissues and Oxidations 42
2.5.1 Skin Tissues 41
2.5.2 Generation of ROS in the Skin 44
2.5.3 Role of Oxidative Stress/ROS in the Skin 47
2.5.3.1 Inflammation 47
2.5.3.2 Increasing Sebum Secretion 48
2.5.3.3 Melanogenesis 48
2.5.3.4 Skin Aging 50
2.6 UV Radiation 54
2.7 Matrix Metalloproteinase (MMP) 56

CHAPTER 3. MATERIALS AND METHODS
3. 1 Experiment Design 64
3.2 Materials 65
3.2.1 Purple-Fleshed Sweet Potato 65
3.2.2 Chemicals 58
3.2.3 Cell Cultures 69
3.3 Purple-Fleshed Sweet Potato Preparation and Extraction 72
3.4 Antioxidant Components 72
3.4.1 Total Monomeric Anthocyanin Content 72
3.4.2 Total Phenolic Content 73
3.4.3 Flavonoids Content 74
3.5 Antioxidant Activity 75
3.5.1 Scavenging Activity on DPPH Radical 75
3.5.2 Ferric-Reducing Antioxidant Power (FRAP) 76
3.5.3 Ferrous Ions Chelating Capacity 76
3.5.4 Scavenging Nitric Oxide 77
3.5.5 Scavenging Hydrogen Peroxide (H2O2) 78
3.6 Anti-aging and Anti-photoaging Activity 78
3.6.1 Effect of TNG 73 Extracts on Fibroblast Cells Viability 80
3.6.2 Population Doubling Level (PDL) 81
3.6.3 X-Gal Cytochemical Assay of SA-βG 81
3.6.4 Assay of SA-βG Activity by FDG Method 82
3.6.5 Determination of Redox Status 84
3.6.5.1 UVB Irradiation 84
3.6.5.2 H2O2 Induced ROS 84
3.6.6 Measurement of Antioxidant Enzymes Activities (SOD) 85
3.6.7 Total Collagen 86
3.6.8 Matrix Metalloproteinase Activity (Gelatin Zymography) 87
3.7 Anticancer Activity (MTT Assay) 88
3.16 Statistical analysis 89
CHAPTER IV. RESULTS
4.1 Purple-Fleshed Sweet Potato Extraction 91
4.2 Antioxidant Components 92
4.2.1 Total Monomeric Anthocyanin (TMA) Content 92
4.2.2 Total Phenolic Content 94
4.2.3 Flavonoids Content 96
4.3 Antioxidant Activity 98
4.3.1 Scavenging Activity on DPPH 98
4.3.2 Ferric-Reducing Antioxidant Power (FRAP) 102
4.3.3 Ferrous Ions Chelating Capacity 104
4.3.4 Scavenging Nitric Oxide 107
4.3.5 Scavenging Hydrogen Peroxide (H2O2) 108
4.4 Anti-aging and Anti-photoaging Activity 111
4.4.1 Effect of TNG 73 Extracts on Cell Viability 111
4.4.2 Population Doubling Level (PDL) 114
4.4.3 Cell Senescence (β-galactoside) in Cultured Human Fibroblasts 115
4.4.4 Determination of Redox Status 118
4.4.4.1 Effect of UVB Irradiation on Cell Proliferation 118
4.4.4.2 Effect of H2O2 on Cell Proliferation 121
4.4.5 Superoxide Dismutase (SOD) Activity 122
4.4.6 Collagen Contents 124
4.4.7 Gelatin Zymography Assay 125
4.4.8 Anticancer Activity 127
CHAPTER V. DISCUSSION

5.1 Purple-Fleshed Sweet Potato Extraction 130
5.2 Antioxidant Components 131
5.2.1 Total Monomeric Anthocyanin (TMA) Content 131
5.2.2 Total Phenolic Content 135
5.2.3 Flavonoids Content 138
5.3 Antioxidant Activity 139
5.3.1 Scavenging Activity on DPPH 139
5.3.2 Ferric-Reducing Antioxidant Power (FRAP) 141
5.3.3 Ferrous Ions Chelating Capacity 142
5.3.4 Scavenging Nitric Oxide 143
5.3.5 Scavenging Hydrogen Peroxide (H2O2) 144
5.4 Anti-aging and Anti-photoaging Activity 146
5.4.1 Effect of TNG 73 Extracts on Fibroblast Cell Viability 146
5.4.2 Population Doubling Level (PDL) 148
5.4.3 Cell Senescence (β-galactoside) in Cultured Human Fibroblasts 149
5.4.4 Effect of UVB Irradiation on Cell Proliferation 151
5.4.5 Effect of H2O2 on Cell Proliferation 154
5.4.6 Superoxide Dismutase (SOD) Activity 156
5.4.7 Collagen Contents 159
5.4.8 Gelatin Zymography Assay 159
5.5 Anticancer Activity 162


CHAPTER VI. CONCLUSION 164
References 167

LIST OF TABLES

Table 1 Range of phenolic compounds in tubers of potato genotypes varying in flesh color 18
Table 2 Relationship between tuber flesh color and anthocyanin content, total carotenoid, antioxidant activity, total phenolics, and oxygen radical absorbance capacity (ORAC) of potato cultivars 28
Table 3 Different purple sweet potato varieties 29

Table 4 Citation Reports of Purple-Fleshed Sweet Potato Studies 34

Table 5 Classification of Matrix Metalloproteinase 61

Table 6 Chemical substances 66

Table 7 Cell Lines 71

Table 8 EC50 of PFSP TNG 73 antioxidant activities 110


LIST OF FIGURES
Figure 1 Sweet Potato Varieties 12
Figure 2 Flavonoids groups 19
Figure 3 Anthocyanidin subclasses 23
Figure 4 Anthocyanins colors based on pH 24
Figure 5 Anthocyanin as a type of flavonoid compound 26
Figure 6 Purple-Fleshed sweet Potato (PFSP) cultivar Tainung 73 31
Figure 7 2000-2013/06 Published PFSP papers per year 33
Figure 8 Cross-section of skin 43
Figure 9 ROS oxidative chain reactions and scavengers 46
Figure 10 Signaling pathways leading to transcription factors activation in response to UV-radiation 53
Figure 11 MMPs function in the tumor microenvironment 60
Figure 12 Experiment Design 64
Figure 13 Tainung 73 specifications 66
Figure 14 Cell Cultures 70
Figure 15 Effects type of solvent and extraction temperature on Total Monomeric Anthocyanin (TMA) content 93
Figure 16 Effects type of solvent and extraction temperature on Total Phenolic content 95
Figure 17 Effects type of solvent and extraction temperature on Flavonoid content 97
Figure 18 DPPH radical-scavenging activity of TNG 73 100
Figure 19 Reducing power of TNG 73 102
Figure 20 Ferrous Chelating Ability of TNG 73 105
Figure 21 Scavenging Activity on Nitric Oxide 108
Figure 22 Scavenging Activity on Hydrogen Peroxide 109
Figure 23 Effects of TNG 73 extract on cell viability 112
Figure 24 Effects of TNG 73 extract on HS-68 population doubling level 114
Figure 25 Permeability of fluorescein produced by FDG and on the intensity of X-Gal staining in HS-68 cells 116
Figure 26 Effect of TNG 73 extract on NIH/3T3 cell viability after UVB irradiation 119
Figure 27 Effect of TNG 73 extract on NIH/3T3 cell viability after H2O2 treatment 121
Figure 28 Effect of TNG 73 extract on NIH/3T3 SOD activity after UVB irradiation and H2O2 treatment 123
Figure 29 Effect of TNG 73 extracts on NIH/3T3 collagen concentration after UVB irradiation 125
Figure 30 MMP expression of NIH/3T3 cells after UVB irradiation by gelatin zymography assay 126
Figure 31 Effects of TNG 73 extract on B16-F10 cells viability 127
Figure 32 Redox reactions 136
Figure 33 MTT formation 147
Figure 34 Principle of SOD assay kit 157

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