Title Page
Contents
ABSTRACT 8
1. Introduction 10
2. Materials and Methods 12
2.1. Materials 12
2.2. Cell Culture 13
2.3. Treatment of Cells with Reagents 14
2.4. Transfection 15
2.5. MitoTracker Staining 16
2.6. Mitochondria-Enriched Fraction 17
2.7. Immunocytochemistry 18
2.8. Detection of Oxidative Phosphorylation 20
2.9. Western Blot 21
2.10. Cellular Reactive Oxygen Species (ROS) Detection 23
2.11. Intracellular ATP Levels Measurement 24
2.12. Animals and Brain Tissue Immunohistochemistry 25
2.13. Data Analysis 26
3. Results 27
3.1. KRGE increased co-expression of GFAP and BVR-A in the fimbria of hippocampus in mice. 27
3.2. Among the HO metabolites, bilirubin contributes particularly to the enhancement of ERRα expression and mitochondrial function. 30
3.3. KRGE induces BVR-A/Bilirubin-mediated LKB1 activation. 33
3.4. KRGE induces LKB1-mediated ERRα, proteins related to mitochondrial function. 36
3.5. Bilirubin increases mitochondrial membrane potential through SIRT1 and 5 activation. 38
3.6. SIRT1 and SIRT5 synergistically regulate KRGE- induced TOM20 and TOM22 expression changes. 40
3.7. Exogenous bilirubin recovers KRGE-induced mitochondrial function improvement in BVR-A deficient astrocytes. 43
3.8. KRGE induces the expression of mitochondrial functional proteins through TOM20/TOM22 in astrocytes. 45
3.9. KRGE enhances mitochondrial functional proteins in mouse brain tissue. 47
4. Discussion 51
5. Conclusion 54
References 55
ABSTRACT (in Korean) 60
Figure 1. KRGE enhances GFAP and BVR-A co-expression in the region of the mouse hippocampal fimbria. 29
Figure 2. Among HO metabolites, bilirubin enhances ERRα expression and mitochondrial function the most. 32
Figure 3. KRGE induces BVR-A/bilirubin-mediated LKB1 phosphorylation. 35
Figure 4. KRGE induces the LKB1-mediated expression enhancement of ERRα and mitochondrial functional proteins. 37
Figure 5. Bilirubin increases mitochondrial membrane potential through SIRT1 and 5 activation. 39
Figure 6. SIRT1 and SIRT5 synergistically regulate KRGE-induced TOM20 and TOM22 expression changes. 41
Figure 7. KRGE-induced cytosolic SIRT1 and mitochondrial SIRT5 increase mitochondrial proteins. 42
Figure 8. Exogenous bilirubin recovers KRGE-induced mitochondrial function improvement in BVR-A-depleted astrocytes. 44
Figure 9. KRGE induces the expression of mitochondrial functional proteins through TOM20/TOM22 in astrocytes. 46
Figure 10. KRGE enhances mitochondrial functional proteins in mouse brain tissue 49
Figure 11. Schemes on the mechanism by which KRGE affects mitochondrial function improvement. 50