Title Page
Contents
Abstract 9
1. Overview 11
1.1. Mitochondria: Definition, Structure, Function 11
1.2. Mitochondrial dysfunction: apoptosis and disease 14
1.3. Research trend of mitochondrial therapy and limitations 17
2. Objective and Strategies 22
3. Introduction 23
3.1. Approaches to mitochondrial therapy for sepsis 23
4. MATERIALS AND METHODS 26
4.1. Mitochondria isolation 26
4.2. Quality control of isolated mitochondria 26
4.3. In vivo cecal slurry model in rats 27
4.4. Mitochondrial respiration in cecal slurry rat model 30
4.5. Reactive oxygen species assay 30
4.6. ATP assay 30
4.7. CFU assay 31
4.8. Lactate assay 31
4.9. Apoptosis study 31
4.10. Mitochondrial dynamics study 32
4.11. Ex vivo splenocyte stimulation with LPS 32
4.12. In vitro LPS stimulation model 33
4.13. Macrophage polarization 37
4.14. Phagocytosis study 37
4.15. Western blot analysis 37
4.16. Cytokine measurements 38
4.17. Distribution of transplanted mitochondria in septic spleen 38
4.18. Statistical anlaysis 39
5. RESULTS 40
5.1. Mitochondrial function after induction of sepsis 40
5.2. Characteristics of isolated mitochondria 40
5.3. Distribution of transplanted mitochondria in septic spleen 43
5.4. Mitochondrial respiration changes in septic spleens 43
5.5. Survival rate, bacterial clearance, and plasma lactate levels 48
5.6. Immunomodulatory effects 48
5.7. In vitro LPS stimulation of human monocytes 52
5.8. Macrophage polarization 56
5.9. Phagocytic effects 56
5.10. Effects on apoptosis in the spleen 59
5.11. Effects on mitochondrial dynamics 59
6. DISCUSSION 62
7. CONCLUSION 66
REFERENCES 67
ABSTRACT IN KOREAN 74
Figure 1. Mitochondrial morphology in animal cells by transmission electron microscopy. 12
Figure 2. Mitochondrial structure in animal cell. 13
Figure 3. Extrinsic and intrinsic apoptosis pathway by ROS. 15
Figure 4. Mitochondrial dysfunction and various diseases. 16
Figure 5. Research trend of mitochondria target materials. 19
Figure 6. Limitation of mitochondrial target anti-oxidants and peptides. 20
Figure 7. Research trend of mitochondrial transplantation. 21
Figure 8. Study strategies for mitochodria therapy. 22
Figure 9. Overview of the effects of mitochondrial transplantation on sepsis and... 25
Figure 10. Conceptual figure of the rat sepsis model. 29
Figure 11. Flow diagram of the experimental procedures used to assess the... 35
Figure 12. Flow diagram of the experimental procedures used to assess the... 36
Figure 13. Respiratory control rates of mitochondria in the rat sepsis model. 41
Figure 14. Characterization of isolated mitochondria from L6 cells. 42
Figure 15. Distributions of transplanted mitochondria in septic spleens. 44
Figure 16. Oxygen consumption trace in muscle, spleen, liver, kidney and heart tissues. 45
Figure 17. Oxygen consumption trace in organ of cecal slurry model in rat. 46
Figure 18. Intracelluar ROS and mitochondria ROS level in septic spleen. 47
Figure 19. Effect of mitochondrial transplantation on sepsis in animal studies. 49
Figure 20. Effect of mitochondrial transplantation on the acute hyperinflammatory... 50
Figure 21. Effect of mitochondrial transplantation on the late immunosuppressive... 51
Figure 22. Effect of mitochondrial transplantation on the in vitro... 53
Figure 23. Effect of mitochondrial transplantation on the in vitro late immuno... 54
Figure 24. Immunomodulatory effect of mitochondria treated with electron... 55
Figure 25. Macrophage polarization in hyperinflammation model. 57
Figure 26. Phagocytosis of E.coli-FITC in immune paralysis model. 58
Figure 27. Effect of mitochondrial transplantation on apoptosis. 60
Figure 28. Effect of Mitochondrial dynamics markers in the spleen were measured... 61