아주대학교

목차

I. INTRODUCTION 1
A. The importance of subcortical white matter injury research 1
B. Animal models of the subcortical ischemic white matter injury 3
1. Animal model induced by vasoconstrictors 4
2. Animal model of chronic cerebrovascular occlusion 5
3. Animal model carrying comorbid vascular risk factors 6
C. Role of BBB integrity in hypertension 8
D. Linking between BBB dysfunction and RAAS 10
E. Aim of the thesis research 13
II. MATERIALS AND METHODS 16
1. Animals 16
2. Surgical procedures for clipping bilateral renal arteries 16
3. Blood pressure measurement 17
4. Magnetic Resonance Imaging (MRI) of the rat brain 18
5. Western blot 19
6. Evaluation of tissue hypoxia using hyproxyprobe 20
7. ELISA analysis 20
8. PCR 21
9. Tissue processing and immunohistochemistry 21
10. Quantitative image analysis 23
11. Preparation of mouse brain endothelial cells 24
12. Measurement of reactive oxygen species generation 24
13. Intracerebroventricular infusion 25
14. Tissue processing and histological assessment 26
15. Behavioral assessment 27
16. Statistical analysis 28
17. Tissue dissection for single cell dissociation of the white matter 28
18. White matter dissociation for single cell sequencing (scRNA-seq) 29
19. Single cell library construction 30
20. Data processing, cell clustering and data analysis 30
III. RESULTS 32
Part A. Establishment of an animal model of white matter injury 32
1. RVH induces progressive white matter hyperintensities 32
2. Pathological alterations in the white matter of RVH rats 41
3. Changes in glial cells in RVH-induced white matter lesions 51
4. Evidence of Tau pathology in RVH-induced white matter injury 55
Part B. Ang II-mediated BBB disruption in RVH model 64
1. BBB disruption in RVH rats 64
2. Potential implication of MMP-9 immunoreactivity in RVH-induce white matter injury 71
3. Increased Ang II in RVH rats 82
4. Effect of the Ang II blocker on brain endothelial cells 85
5. Effect of the Ang II blocker in RVHT rat 88
Part C. Single cell RNA sequencing analysis of glial transcriptomics in RVH model 97
1. Clustering of different cell type in the white matter 97
2. Cell-type specific analysis of DEGs in RVH-induced white matter injury 105
3. Identifying glial subtypes with differing functions 110
V. SUMMARY AND CONCLUSION 129
REFERENCE 130
국문 요약 144