Title Page
Contents
Background 13
Part Ⅰ. Improved lactate metabolism and endurance performance in mice treated with Ginseng berry extract. 16
1.1. Abstract 17
1.2. Introduction 19
1.3. Abbreviations 23
1.4. Materials and methods 24
1.4.1. Animals 24
1.4.2. Preparation of GBP (Ginseng berry polysaccharide) 24
1.4.3. GBP (Ginseng berry polysaccharide) administration 25
1.4.4. Exercise performance 25
1.4.5. Cell culture 26
1.4.6. Measurement of OCR (Oxygen Consumption Rate) 26
1.4.7. Cryosections, Staining analysis 26
1.4.8. Measurement of Cross section area (CSA) 27
1.4.9. Blood chemistry analysis 27
1.4.10. Enzyme-linked immunosorbent assay (ELISA) 27
1.4.11. RNA Isolation and cDNA synthesis 28
1.4.12. qRT-PCR 28
1.4.13. Western blot 28
1.4.14. RNA sequencing and gene set enrichment analysis 29
1.4.15. Liquid chromatography-tandem mass spectrometry 29
1.4.16. Sample preparation for NAD⁺ analysis in muscle tissue by LC-MS/MS 30
1.4.17. Statistical analysis 30
1.5. Results 31
1.5.1. Ginseng berry polysaccharide increased the oxygen consumption rate. 31
1.5.2. Gene set enrichment analysis (GSEA) demonstrates Ginseng berry polysaccharide induces mitochondrial gene expression in C2C12 cells. 32
1.5.3. Gene set enrichment analysis confirms the effect of Re on mitochondria and age-associated pathways. 32
1.5.4. Ginseng berry polysaccharide administered animal model experiments. 33
1.5.5. Body weight, Food intake, and tissue mass of Ginseng berry polysaccharide administered animal model. 34
1.5.6. Ginseng berry polysaccharide enhanced exercise performance ability and clearly reduced lactate levels. 34
1.5.7. The phenotype of Ginseng berry polysaccharide administered animal model. 35
1.5.8. Biological effects of Ginseng berry polysaccharide 35
1.5.9. The NAD⁺ level increases by administered Ginseng berry polysaccharide. 36
1.5.10. The AMP-activated protein kinase (AMPK) activity levels increase by administered Ginseng berry polysaccharide. 38
1.6. Discussion 40
1.8. References 65
Part Ⅱ. Improved gut microbiota composition and muscle function in a mice model of cancer cachexia 70
2.1. Abstract 71
2.2. Introduction 73
2.3. Abbreviations 76
2.4. Materials and methods 77
2.4.1. Cancer cachexia models. 77
2.4.2. Preparation of microbiota 77
2.4.3. Grip strength 77
2.4.4. Cell culture 78
2.4.5. Tissue preparations, Histology 78
2.4.6. Collection of fecal samples 78
2.4.7. Analysis of composition the microbiota 79
2.4.8. Analysis of metagenomic 79
2.4.9. Analysis of targeted metabolites 79
2.4.10. Statistical analysis 80
2.5. Results 81
2.5.1. Phenotype of the cancer cachexia mice model compared with normal mice. 81
2.5.2. Cancer cachexia models show decreased muscle strength and cross section area. 82
2.5.3. Altered composition of gut microbiome in cancer cachexia mice model. 82
2.5.4. Differential abundance analysis of gut microbiota by DESeq2. 82
2.5.5. Concentration and molar ratio of gut microbial metabolites (SCFA) detected by GC-FID in mouse fecal samples. 83
2.5.6. Phenotype of Lactiplantibacillus plantarum administered cancer cachexia model. 84
2.5.7. Biological effects of Lactiplantibacillus plantarum administered cancer cachexia model. 85
2.6. Discussion 86
2.8. References 107
논문요약 111
Part Ⅰ. Improved lactate metabolism and endurance performance in mice treated with Ginseng berry extract 8
Figure 1. GBP increased the oxygen consumption rate. 45
Figure 2. Gene set enrichment analysis (GSEA) demonstrates GBP induces mitochondrial gene expression in C2C12 cells. 47
Figure 3. Gene set enrichment analysis confirms the effect of Re on mitochondria and age- associated pathways. 49
Figure 4. GBP-administered animal model experiments protocol. 51
Figure 5. Graph of Body weight and Food intake of the GBP-administered animal model. 52
Figure 5. Graph comparing the mass of tissue in the GBP-administered group or Control group. 54
Figure 6. GBP enhanced exercise performance ability and clearly reduced lactate levels. 57
Figure 7. The phenotype of GBP-administered animal model. 59
Figure 8. Biological effects of Ginseng berry polysaccharide. 61
Figure 9. The NAD⁺ Level increases by administered GBP. 62
Figure 10. The AMP-activated protein kinase (AMPK) activity levels increase by administered Ginseng berry polysaccharide. 64
Part Ⅱ. Improved gut microbiota composition and muscle function in a mice model of cancer cachexia 11
Figure 1. Phenotype of the cancer cachexia mice model compared with normal mice 91
Figure 2. Cancer cachexia models show decreased muscle strength and cross section area. 94
Figure 3. Altered composition of Gut microbiome in cancer cachexia mice model. 96
Figure 4. Differential abundance analysis of gut microbiota by DESeq2 97
Figure 5. Concentration and molar ratio of gut microbial metabolites (SCFA) detected by GC- FID in mouse fecal samples. 99
Figure 6. Muscle and tissue phenotype of Lactiplantibacillus plantarum administered cancer cachexia model. 102
Figure 7. Analysis muscle strength of Lactiplantibacillus plantarum administered cancer cachexia model. 103
Figure 8. Evaluation of the Cross-section area of Lactiplantibacillus plantarum administered cancer cachexia model. 105
Figure 9. The effect of Lactiplantibacillus plantarum administration in cancer cachexia models. 106