Title Page
Contents
ABSTRACT 8
1. Introduction 10
2. Materials and Methods 12
2.1. Materials 12
2.2. Animals 13
2.3. Brain Tissue Immunohistochemistry 14
2.4. Cell Culture 15
2.5. Transfection 16
2.6. Immunocytochemistry 17
2.7. MitoTracker Staining 18
2.8. Western Blot 19
2.9. Oxidative Phosphorylation (OXPHOS) Detection 21
2.10. Oxygen Consumption Assay 22
2.11. Intracellular NAD⁺/NADH Measurement 23
2.12. Mitochondria-Cytosol Fraction 24
2.13. Intracellular ATP levels 25
2.14. Protein Intensity in GFAP⁺-Astrocytes 26
2.15. Data Evaluation 27
3. Results 28
3.1. VEGF induced by KRGE in astrocytes around the corpus callosum. 28
3.2. KRGE stimulates HIF-1α protein levels around the corpus callosum. 30
3.3. SIRT3, induced by KRGE, regulates HIF-1α protein levels. 33
3.4. Nampt, induced by KRGE, did not increase NAD⁺/NADH ratio or HIF-1α protein levels. 36
3.5. Influence of HIF-1α on mitochondrial proteins and import mechanisms in KRGE-treated astrocytes. 38
3.6. Impact of KRGE-induced SIRT3 on astrocytic mitochondrial functions. 41
3.7. KRGE upregulated HIF-1α expression in astrocytes through increasing Tom22-mediated oxygen consumption. 44
3.8. KRGE enhances SIRT3 and Tom22 to stabilize HIF-1α protein via PHD2. 47
4. Discussion 50
5. Conclusion 54
References 55
Abstract (in Korean) 61
Figure 1. VEGF induced by KRGE in astrocytes around the corpus callosum. 29
Figure 2. KRGE (0.25 mg / mL) induced HIF-1α in astrocytes above the corpus callosum. 32
Figure 3. SIRT3, induced by KRGE, regulates HIF-1α protein levels. 35
Figure 4. The effect of KRGE-induced Nampt on NAD⁺/NADH ratio and HIF-1 α protein levels. 37
Figure 5. Impact of HIF-1α on Mitochondria, Tom20, and Tom22 expression in KRGE-treated in vitro astrocytes. 40
Figure 6. SIRT3, induced by KRGE, regulates astrocytic mitochondrial functions. 43
Figure 7. KRGE enhances HIF-1α in in vitro astrocytes by boosting Tom22- mediated oxygen consumption. 46
Figure 8. KRGE-mediated SIRT3 and Tom22 maintain HIF-1α protein via PHD2. 48
Figure 9. Schemes on the molecular mechanisms of this study. 49