목차

ABSTRACT ⅰ
CONTENTS ⅲ
Introduction 1
1.1 Increased oxidative stress from reactive oxygen species (ROS) imbalance 1
1.2 Antioxidants that reduce oxidative stress 3
1.3 In vitro and in vivo Experiments to evaluate antioxidant activity 5
1.4 Seaweed polysaccharides used in the development of natural antioxidants 6
1.5 Extracting seaweed polysaccharides using carbohydrate enzymes 9
Materials and Methods 11
2.1 Chemicals and reagents 11
2.2 Seaweeds materials and extractions 12
2.3 Yield 12
2.4 Fractionation of crude polysaccharide (CPS) 13
2.5 Analysis of sugar composition 14
2.6 Molecular weight analysis 14
2.7 Hydrogen peroxide scavenging assay 15
2.8 Cell culture 16
2.9 Intracellular ROS scavenging assay using DCF-DA 16
2.10 Assessment of cell viability 17
2.11 Detection of apoptosis by Hoechst 33342 and propidium iodide (PI) staining and flow cytometry 17
2.12 Western blot assay 19
2.13 Zebrafish management 20
2.14 Protection effect of CPS extracts against intracellular ROS and cell death induced by H2O2 in zebrafish
model 21
2.15 Statistical analysis 22
Results 23
3.1 Yield and DPPH H2O2 scavenging activity of seaweeds extracts 23
3.2 Protective effect of seaweed extracts against oxidative stress induced by H2O2 in vitro 25
3.3.1 Extraction yield and DPPH and H2O2 scavenging activity of C. sinuosa enzyme extracts 28
3.3.2 Protective effect of C. sinuosa enzyme extracts against in vitro oxidative stress induced by H2O2 30
3.3.3 Protective effect of the CSU in in vitro cell death and DNA fragmentation induced by oxidative stress 33
3.3.4 Effect of CSU on expression levels of apoptosis related Proteins 36
3.3.5 Flow cytometry analysis of the protective effect of the CSU against apoptosis 38
3.3.6 Yield and DPPH and H2O2 scavenging activity of CSU and CPS fractions 40
3.3.7 Protective effect of CSU CPS fractions against in vitro oxidative stress induced by H2O2 42
3.3.8 Protective effect of the CPS1 in in vitro cell death and DNA fragmentation induced by oxidative stress 44
3.3.9 Effect of CPS1 on expression levels of apoptosis related Proteins 46
3.3.10 Molecular weight analysis of C. sinuosa extract 48
3.3.11 Sugar composition of C. sinuosa extracts 50
3.4.1 Extraction yield and DPPH, H2O2-scavenging activity of G. amansii enzyme extracts 52
3.4.2 Protective effect of G. amansii enzyme extracts against oxidative stress induced by H2O2 in vitro 54
3.4.3 Protective effect of the GAT in increasing cell death and DNA fragmentation induced by oxidative stress
in vitro 56
3.4.4 Effect of GAT on expression levels of apoptosis related Proteins 58
3.4.5 Flow cytometry analysis of the protective effect of the GAT against apoptosis 60
3.4.6 Yield and DPPH and H2O2 scavenging activity of GAT and CPS fractions 62
3.4.7 Protective effect of GAT CPS fractions against in vitro oxidative stress induced by H2O2 64
3.4.8 Protective effect of the CPS4 in increasing cell death and DNA fragmentation induced by oxidative stress
in vitro 66
3.4.9 Effect of CPS4 on expression levels of apoptosis related proteins 69
3.4.10 Protective effect of the CPS4 fraction against oxidative stress induced by H2O2 in zebrafish embryos
71
3.4.11 Molecular weight analysis of G.amansii Extracts 74
3.4.12 Sugar composition of G.amansii extracts 76
Discussion 78
Reference 84
국문초록 91