목차

INTRODUCTION 1
REFERENCES 5
CHAPTER 1. Antioxidative and neuroprotective effects of Carthamus tinctorius L. seed and Taraxacum coreanum in vitro and cellular system 11
Study 1. Co-treatment of Carthamus tinctorius L. seed and Taraxacum coreanum attenuates H2O2-induced oxidative stress and apoptosis in SH-SY5Y cells 13
Abstract 13
1. Introduction 14
2. Materials and Methods 15
2.1. Materials 15
2.2. 1,1-Dephenyl-2-picrylhydrazyl (DPPH) scavenging assay 16
2.3. Hydroxyl radical (OH) scavenging assay 16
2.4. Cell culture and treatment 17
2.5. MTT assay 17
2.6. DCF-DA assay 17
2.7. Lactate dehydrogenase (LDH) assay 18
2.8. Nitric oxide (NO) assay 18
2.9. Hoechst 33342 staining 18
2.10. Western blot analysis 18
2.11. Statistical analysis 19
3. Results 19
3.1. DPPH and OH radical scavenging activities 19
3.2. Protective effect from H2O2-induced neurotoxicity in SH-SY5Y cells 19
3.3. Protective effect on H2O2-induced ROS production in SH-SY5Y cells 23
3.4. Effect on cell apoptosis H2O2-treated SH-SY5Y cells 23
3.5. Effect on expression of antioxidative enzymes in H2O2-treated SH-SY5Y cells 27
4. Discussion 27
5. References 32
Study 2. Co-treatment with Carthamus tinctorius L. seed and Taraxacum coreanum synergistically suppresses Aβ25-35-induced neurotoxicity by altering APP processing 37
Abstract 37
1. Introduction 37
2. Materials and Methods 39
2.1. Reagents 39
2.2. Sample preparation 39
2.3. Aβ25-35 stocks 39
2.4. Cell culture and treatment 40
2.5. Viability and ROS assays 40
2.6. Western blotting 40
2.7. Statistical analysis 41
3. Results 41
3.1. Effect on cell viability in Aβ25-35-treated SH-SY5Y cells 41
3.2. Synergistic effect of ROS inhibition in Aβ25-35-treated SH-SY5Y cells 41
3.3. Synergistic anti-amyloidogenic effect in Aβ25-35-treated SH-SY5Y cells 44
3.4. Synergistic anti-apoptotic effect in Aβ25-35-treated SH-SY5Y cells 44
4. Discussion 47
5. References 48
CHAPTER 2. Determination of Carthamus tinctorius L. seed and Taraxacum coreanum at different ratios on synergistic effect from cognitive impairment in vivo 53
Study 1. Combination of Carthamus tinctorius L. seed and Taraxacum coreanum exerts synergistic exerts synergistic effects on learning and memory function by regulating metabolism of amyloid beta in mice 55
Abstract 55
1. Introduction 55
2. Materials and Methods 57
2.1. Reagents 57
2.2. Sample preparation 58
2.3. Animals and experimental design 58
2.4. Aβ25-35-infused mouse model 59
2.5. T-maze test 59
2.6. Novel objective recognition test 59
2.7. Morris water maze test 61
2.8. Western blot analysis 62
2.9. Statistical analysis 62
3. Results 62
3.1. Effects of the combination at different ratios on body weight, organ weight and liver function in Aβ-infused mice 62
3.2. Synergistic effects of the combination at different ratios on the T-maze test in Aβ-infused mice 66
3.3. Synergistic effects of the combination at different ratios on the novel object recognition test in Aβ-infused mice 66
3.4. Synergistic effects of combination at different ratios on the Morris water maze test in Aβ-infused mice 69
3.5. Synergistic effects of the combination at different ratios on amyloidogenic pathway 71
3.6. Synergistic effect of the combination at different ratios on Aβ clearance and degradation 71
4. Discussion 75
5. References 81
Study 2. Synergistic protection by co-administration of Carthamus tinctorius L. seed and Taraxacum coreanum from memory impairment and inflammation on scopolamine-injected mice 91
Abstract 91
1. Introduction 91
2. Materials and Methods 93
2.1. Sample preparation 93
2.2. HPLC analyses 93
2.3. Animals 94
2.4. Experimental design 94
2.5. T-maze test 98
2.6. Novel object recognition test 98
2.7. Biochemical analysis of blood 99
2.8. Western blot analysis 99
2.9. Statistical analysis 100
3. Results 100
3.1. Chromatographic analysis of C. tinctorius L. seed and T. coreanum 100
3.2. Serum biochemistry in scopolamine-injected mouse 100
3.3. Effect on spatial memory ability in scopolamine-injected mouse 101
3.4. Effect on novel object cognitive ability in scopolamine-injected mouse 101
3.5. Effect on iNOS, COX-2, and GFAP protein expression in scopolamine-injected mouse 102
3.6. Effect on AChE and BDNF protein expression in scopolamine-injected mouse 102
4. Discussion 107
5. References 114
CHAPTER 3. Effect of combination with Carthamus tinctorius L. seed and Taraxacum coreanum at different concentration on cognitive improvement in vivo 121
Study 1. Herbal mixture of Carthamus tinctorius L. seed and Taraxacum coreanum attenuates amyloid beta-induced cognitive dysfunction in vivo 123
Abstract 123
1. Introduction 123
2. Materials and Methods 125
2.1. Sample preparation 125
2.2. Aβ peptide preparation and injection 126
2.3. Animals and treatments 126
2.4. T-maze test 127
2.5. Novel object recognition test 127
2.6. Morris water maze test 129
2.7. Tests for aspartate transaminase (AST) and alanine transaminase (ALT) 129
2.8. Western blotting 130
2.9. Statistical analysis 130
3. Results 130
3.1. Effect of CT on body weight change and liver function in Aβ25-35-infused mice 130
3.2. Improvement of Aβ25-35-induced spatial impairment in T-maze test in CT-treated mice 131
3.3. Improvement of Aβ25-35-induced object recognition impairment in novel object recognition test in CT-treated mice 131
3.4. Improvement of Aβ25-35-induced long-term spatial memory impairment in Morris water maze test in CT-treated mice 136
3.5.Regulatory effect of CT on amyloidogenic pathway in Aβ25-35-infused mice 136
4. Discussion 137
5. References 143
Study 2. Protective effects of combination with Carthamus tinctorius L. seed and Taraxacum coreanum on scopolamine-induced memory impairment in mice 151
Abstract 151
1. Introduction 151
2. Materials and Methods 153
2.1. Sample preparation 153
2.2. Experimental design 154
2.3. Novel object recognition test 155
2.4. Passive avoidance test 155
2.5. T-maze test 156
2.6. Morris water maze test 156
2.7. Western blot analysis 157
2.8. Statistical analysis 157
3. Results 158
3.1. Effect on object recognition impairment in novel object recognition test 158
3.2. Effect on working memory deficit in passive avoidance test 158
3.3. Effect on spatial impairment in T-maze test 158
3.4. Effect on learning and memory impairment in Morris water maze test 161
3.5. Effect on protein expressions of acetylcholinesterase and butyrylcholinesterase 161
3.6. Effect on protein expressions of BDNF and p-CREB 165
3.7. Effect on amyloid precursor protein expression 165
4. Discussion 165
5. References 170
SUMMARY AND CONCLUSION 177
ABSTRACT (In Korean) 179