Title Page
ABSTRACT
ABSTRACT IN KOREAN
PREFACE
Contents
LIST OF ABBREVIATIONS 25
Background 29
1. Oxidative stress 30
2. Alcohol-induced oxidative stress 33
3. Natural substances that help with hangover relief 38
4. Hangover-relief effect of ginseng 42
5. Application of the ginseng berry 43
Chapter 1. Fermentation Characteristics of Ginseng Berry by Saccharomyces cerevisiae Isolated from Sugar-preserved Ginseng 49
Abstract 50
1-1. Introduction 52
1-2. Materials and Methods 55
1-2-1. Strain isolation 55
1-2-2. Strain identification 55
1-2-3. Ginseng berry fermentation 56
1-2-4. Analysis of component changes during fermentation 58
1-2-5. Analysis of ginsenosides during fermentation 58
1-2-6. Assay of radical scavenging acidity during fermentation 59
1-2-7. Intestinal permeability using an intestinal sac 60
1-2-8. Statistical analysis 61
1-3. Results 62
1-3-1. Isolation and identification of β-glucosidase producing microbes 62
1-3-2. Changes in components during ginseng berry fermentation 67
1-3-3. Changes in ginsenosides of fermented extracts during fermentation 69
1-3-4. Evaluation of intestinal absorption rate of fermented ginseng berry using the intestinal sac 72
1-4. Discussion 76
1-5. Conclusion 80
Chapter 2. Fermentation Characteristics and Radical Scavenging Capacities of Ginseng Berry Kombucha Fermented by Saccharomyces cerevisiae and Gluconobacter oxydans 81
Abstract 82
2-1. Introduction 84
2-2. Materials and Methods 86
2-2-1. Strains 86
2-2-2. Fermentation of ginseng berry kombucha (GBK) 86
2-2-3. Analysis of component changes during fermentation 88
2-2-4. Microbial analysis during fermentation of GBK 88
2-2-5. Estimation of radical scavenging activity during fermentation 89
2-2-6. Analysis of flavonoid and ginsenoside composition in GBK extracts 89
2-2-7. Estimation of survival rate in Drosophila model 90
2-2-8. Oxidation level in Drosophila (mRNA expression of related oxidative factors) 91
2-2-9. Statistical analysis 92
2-3. Results 93
2-3-1. Component changes during the fermentation of ginseng berry kombucha (GBK) 93
2-3-2. Changes in radical scavenging activity during fermentation 97
2-3-3. Changes in flavonoid and ginsenoside contents before and after fermentation 100
2-3-4. Effects of ginseng berry kombucha (GBK) on survival rate and relief of oxidative stress in male fruit flies under oxidative stress conditions 103
2-4. Discussion 106
2-5. Conclusion 111
Chapter 3. Hangover-relieving Effect of Ginseng Berry Kombucha Fermented by Saccharomyces cerevisiae and Gluconobacter oxydans in Ethanol-treated Cells and Mice model 112
Abstract 113
3-1. Introduction 114
3-2. Materials and methods 118
3-2-1. GBK preparation 118
3-2-2. Analysis of radical scavenging activity of fermented GBK 118
3-2-3. Analysis of the content of ginsenosides in GBK 119
3-2-4. Cell culture 119
3-2-5. Cell viability assay 119
3-2-6. mRNA expression analysis of gene related to oxidative stress and alcohol metabolism 120
3-2-7. Malondialdehyde (MDA) assay 121
3-2-8. Animals 122
3-2-9. Analysis of blood ethanol and acetaldehyde concentrations 123
3-2-10. Analysis of ADH and ALDH activities in the liver tissue 123
3-2-11. Analysis of serum AST, ALT, Glucose, and LDH levels 123
3-2-12. Behavioral analysis following ethanol administration 124
3-2-13. Statistical analysis 126
3-3. Results 127
3-3-1. Fermentation-mediated changes in radical scavenging activity and ginsenoside content 127
3-3-2. Effects of GBK on the viability of ethanol-treated HepG2 cells 130
3-3-3. Effects of GBK on oxidative stress elimination and alcohol metabolism-related gene expression in ethanol-treated HepG2 cells 132
3-3-4. Effects of GBK on oxidative stress elimination-related protein expression in ethanol-treated HepG2 cells 137
3-3-5. Effects of GBK on ethanol and acetaldehyde concentrations in the blood of ethanol-treated mice 139
3-3-6. Effects of GBK on ADH and ALDH activities in the liver of ethanol-treated mice 141
3-3-7. Effects of GBK on AST, ALT, and LDH levels in the blood of ethanol-treated mice 143
3-3-8. Effects of GBK on ethanol-induced behavior 145
3-4. Discussion 147
3-5. Conclusion 152
Conclusion 153
References 156
Table 1-1. Amount and contents of ginsenoside in ginseng berry, leaf and root part 47
Table 1-2. Carbohydrate utilization characteristics of isolated LB-5 strain 66
Table 1-3. Changes in ginsenosides from alcoholic extracts of fermented products during ginseng berry fermentation. 70
Table 1-4. Intestinal permeation amount of ginsenosides before and after ginseng berry fermentation in the by intestinal sac 75
Table 2-1. Content of different microorganisms during the fermentation of ginseng berry kombucha (GBK) 96
Table 2-2. Changes in content of flavonoids before and after ginseng berry kombucha (GBK) fermentation 101
Table 2-3. Changes in content of ginsenosides before and after ginseng berry kombucha (GBK) fermentation 102
Table 3-1. Content of ginsenosides of ginseng berry kombucha (GBK) 129
Table 3-2. Effects of ginseng berry kombucha on serum AST, ALT, GLU, and LDH levels in mice. 144
Figure 1-1. Schematic figure of the link between ROS, oxidative stress and their effects on the human body. Oxidative stress is the imbalnce that oxxurs when... 32
Figure 1-2. The metabolism of beverage alcohol (i.e., ehtanol) by the alcohol dehydrogenase (ADH) pathway. ADH=alcohol dehydrogenase;... 35
Figure 1-3. Oxidative stress and the inflammatory response to alcohol consumption. Arrows in red represent the relationship between alcohol... 36
Figure 1-4. Natural products for the prevention and treatment of hangover and alcohol use disorder 41
Figure 1-5. The different constitutions and contents of ginseng berry and root. Gingseng berry comprises approximately 20% ginsenoside compared to... 46
Figure 1-6. Ginsenoside transformation pathways during biological processing. The sugar moieties linked to C–3, C–6, or C–20 in the major ginsenosides are... 48
Figure 1-7. Schematic of ginseng berry fermentation and extraction 57
Figure 1-8. Colonies of β-glucosidase-producing strains from sugar-preserved ginseng in MRS (A) and PDA (B) esculin medium. MRS, de Man-Rogosa-... 64
Figure 1-9. Phylogenetic tree based on the 18S rRNA gene sequence of M-5 strain isolated from sugar-preserved ginseng. 65
Figure 1-10. Changes in general components (reducing sugar, protein, polyphenols contents, and pH) and cell number (A) and radical scavenging... 68
Figure 1-11. Intestinal permeation amount of polyphenols (A) and polyphenol compounds (B) before and after ginseng berry fermentation in the intestinal sac.... 74
Figure 2-1. Schematic of ginseng berry kombucha fermentation and extraction 87
Figure 2-2. Changes of titratable acidity and pH (A), and components (B) during the fermentation of ginseng berry kombucha (GBK)... 95
Figure 2-3. Changes in ABTS and DPPH radical scavenging activities during fermentation of ginseng berry kombucha (GBK)... 98
Figure 2-4. IC 50 value on ABTS (A) and DPPH radical (B) and FRAP value (C) before and after fermentation of ginseng berry kombucha (GBK)... 99
Figure 2-5. Effects of ginseng berry kombucha (GBK) on the survival rate in Drosophila treated with 15% H₂O₂... 104
Figure 2-6. Effects of ginseng berry kombucha (GBK) on ROS production (A) and antioxidant enzyme-related mRNA expression (B-D) in Drosophila treated... 105
Figure 3-1. Schematics illustrating the apparatus of the balance beam test (A) and elevated plus maze test (B). 125
Figure 3-2. IC₅₀ values for ABTS and DPPH radicals during the fermentat ion of ginseng berry kombucha (GBK). Values are mean ± standard devia... 128
Figure 3-3. Effects of ethanol alone (A) and GBK and ethanol mixed treatment (B) on the viability of HepG2 cells. HepG2 cells were treated with GBK in the... 131
Figure 3-4. Effects of GBK on malondialdehyde content (A) and mRNA expression of factors related to oxidative stress (B-F) in ethanol-treated HepG2 cells. HepG2 cells were treated with GBK in the presence or absence of ethanol for 24 h. Values are... 134
Figure 3-5. Effects of GBK on alcohol metabolism (A-C) in ethanol-treated HepG2 cells. HepG2 cells were treated with GBK in the presence or absence of ethanol for 24 h. Values are mean ± SD. *p 〈0.05 and ***p 〈0.001 vs. NOR group; #p 〈0.05, ##p...[이미지참조] 136
Figure 3-6. Effects of GBK on the protein expression of Nrf2 (A), Keap1 (B), and HO-1 (C) in ethanol-treated HepG2 cells. HepG2 cells were treated with GBK in the presence or absence of ethanol for 24 h. Values are mean ± SD. ***p 〈0.001 vs. NOR... 138
Figure 3-7. Effects of GBK on serum ethanol (A) and acetaldehyde (B) levels in mice. Values are mean ± standard of the mean (SEM). GBK was... 140
Figure 3-8. Effects of GBK on ADH activity (A) and ALDH activity (B) in mouse liver tissues. Values are mean ± SEM. GBK was administered orally 30... 142
Figure 3-9. Effects of GBK on ethanol-induced behavioral changes in mice. The balance beam test (A, B) and elevated plus maze test (C). Values are mean ± SEM. GBK was administered orally 30 min before ethanol administration. ##p 〈0.01 and ###p...[이미지참조] 146