Synthesis and Antioxidant Activity Evaluation of Oligomeric Anthocyanin, using the Anthocyanin Extracted from Grape Skins : 황진우 [지음]

다국어 초록 (Multilingual Abstract)

Anthocyanins effectively neutralize free radicals by the antioxidative and anti-aging, and dementia prevention have been reported. In addition, anthocyanins reproduction of rhodopsin by facilitating night vision impairment, blurred vision, eye fatigue due to physical and mental fatigue, and lack of effective rhodopsin effect is due to various eye diseases. In addition, oligomeric anthocyanins are well known to higher effect than anthocyanin.
In this study, we found that the production of enzyme using A. niger for the synthesis of Oligomeric anthocyanin. The present studing also demonstrated to determine the synthesis of the oligomeric anthocyanin by HPLC and ESI-MS. The present studing also demonstrated to determine the yield of a synthesized oligomeric anthocyanin. As a result, the glucosidase showed high efficiency of synthesis of oligomeric anthocyanin.
The free radical scavenging activity of oligomeric anthocyanins was evaluated for hydroxyl radical by using the electron spin resonance (ESR) spectroscopy. The hydroxyl radical scavenging activity of oligomeric anthocyanins was increased in a dose-dependent manner, and the 50% of inhibitory concentration (IC50) values were 0.454, 0.283, 0.217 and 0.279 mg/ml, respectively.
Oligomeric anthocyanins enhanced the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione-S-transferase (GST) in ARPE-19 cells. In addition, Oligomeric anthocyanins inhibited the t-BHP-induced G2/M phase arrest, by reducing the expression of p21 and inducing the expression of cyclin B.
It confirmed the stability of oligomer-rich anthocyanins through reverse mutation test, in vivo micronucleus test in mouse, single-dose oral toxicity study in SD rats and Beagle dogs, 4 weeks repeated oral dose toxicity studies in SD rats
Taken all together, the present results demonstrates in details of oligomeric anthocyanins have potent antioxidant activity and stability.

목차 (Table of Contents)

• List of Tables iii
• List of Scheme iv
• List of Figures iv
• Abstract viii
• Chapter 1. Introduction 1
• 1.1. Anthocyanin? 1
• 1.2. Functions of anthocyanins 4
• 1.3. Biological activities of anthocyanin 4
• 1.4. Antioxidant properties and free radical scavenging properties 8
• 1.5. Synthesis of anthocyanin 12
• 1.6. Anthocyanin effect in eye 17
• 1.7. Oxidative damage in the hyman body 19
• Chapter 2. Materials and Methods 24
• 2.1. Materials 24
• 2.2. Culture medium of Aspergillus niger 24
• 2.3. Synthesis of oligomer-rich anthocyanin by fermentation using Aspergillus niger 24
• 2.4. Separation of fermented crude enzyme from the culture of Aspergillus niger 25
• 2.5. Synthesis of oligomer-rich anthocyanin using crude enzyme 25
• 2.6. Analysis of crude enzyme from the culture of Aspergillus niger 29
• 2.7. Synthesis of oligomer-rich anthocyanin using various enzyme 29
• 2.8. Hydroxyl radical scavenging activity by an ESR spectrometer 29
• 2.9. Cell culture 31
• 2.10. Cytotoxicity assay in vitro 31
• 2.11. Determination of antioxidative enzyme activity 32
• 2.11.1. Superoxide dismutase (SOD) assay 32
• 2.11.2. Catalase (CAT) assay 32
• 2.11.3. Activity assay of glutathione peroxidase (GPx) 32
• 2.11.4. Glutathione-S-transferase (GST) assay 33
• 2.12. Flow cytomerter analysis 33
• 2.13. Immuno blot analysis 33
• 2.14. Stability testing of oligomer-rich anthocyanin using glucosidase 34
• 2.15. Statistical analysis 34
• Chapter 3. Results and Discussion 35
• 3.1. Synthesis of oligomer-rich anthocyanin 35
• 3.2. Antioxidant activity of oligomer-rich anthocyanin 64
• 3.2.1. Hydroxyl radical scavenging activity of oligomer-rich anthocyanins 64
• 3.2.2. Cytotoxicity of oligomer-rich anthocyanins 64
• 3.2.3. Antioxidative enzyme activity of oligomer0rich anthocyanins on retinal cells 73
• 3.2.3.1. Superoxide dismutase activity 73
• 3.2.3.2. Catalase activity 73
• 3.2.3.3. Glutathione peroxidase activity 77
• 3.2.3.4. Glutathione-S-transferase activity 77
• 3.2.4. Protective effect of oligomer-rich anthocyanins on retinal cells 80
• 3.2.5. Changes of G2/M phase-related gene expression 80
• 3.3. Stability testing of oligomer-rich anthocyanin 84
• 3.3.1. Reverse mutation test 84
• 3.3.2. In vivo mcronucleus test in mouse 93
• 3.3.3. Single-dose oral toxicity study of oligomer-rich anthocyanin in Sprague-Dawley rats 99
• 3.3.4. 4 week repeated oral dose toxicity studies with oligomer-rich anthocyanin in Sprague-Dawley rats 99
• 3.3.5. Single-dose oral toxicity study of oligomer-rich anthocyanin in Beagle dogs 104
• Chapter 4. Conclusions 119
• References 120
• Abstract (in Korean) 139