퇴행성 뇌질환에서 Mesenchymal stem cells의 영향
Neuroprotective effect of mesenchymal stem cells in neurodegenerative disease : focusing on Parkinsonian disease
- 주제(키워드) mesenchymal stem cell , multiple system atrophy , blood brain barrier.
- 발행기관 아주대학교
- 지도교수 이필휴, 안영환
- 발행년도 2010
- 학위수여년월 2010. 8
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000010968
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Mesenchymal stem cells (MSCs) are themselves capable of multi-potency, with differentiation under appropriate conditions into chondrocytes, skeletal myocytes, and neurons. MSCs have been also known to pose neuroprotective effects through secreting various cytotrophic factors. Based on our previous studies demonstrating neuroprotective effect of hMSCs in Parkinson’s disease, we extended our investigations into other parkinsonian disease in addition to properties of hMSCs on blood-brain barrier. . PART A. Human mesenchymal stem cells exerts neuroprotection in an animal model of double lesion-induced multiple system atrophy-parkinsonism (MSA-P). PART B. Inhibition of blood brain barrier (BBB) permeability by hMSCs. First, the double lesion-induced MSA-P was established with co-injections of MPTP (10mg/day, total dose 90 mg/kg for 9 days) and 3- NP (total dose 450 mg/kg for 9 days, 12hr interval). At one day after last injection, hMSCs were injected into the tail vain (1X106cells/ml). Three groups of mice were compared (control group, only MPTP+3-NP group, hMSC treatment in MPTP+3-NP group) through histopathological and behavioral analysis. Compared to only MPTP+3-NP-treated mice, hMSCs treatment in double lesion mice significantly increased survival of TH- and NeuN-immunoreactive cells in the substantia nigra and the striatum, respectively. Additionally, hMSC treatment significantly decreased Iba-1, GFAP and increased Calbindine immunoreactive cells in the substantia nigra and the striatum. Behavioral analysis showed that the decent times on the top of a vertical wooden pole was significantly decreased in hMSCs-treated double lesion mice, comparable to controls. This study demonstrates that hMSCs treatment had a protective effect on loss of neurons in the substantia nigra and the striatum induced by MPTP+3-NP through a variety of mechanisms, such as anti-inflammatory actions, anti-apoptotic effect. Second, the neuroinflammation plays collectively suggest that excessive neutrophil infiltration and environmental factors, such as lower astrocyte density and higher BBB permeability, contribute to severe inflammation and neuronal death in the SNpc. At 4hour (4hr) after LPS injection, hMSCs were injected into the tail vain (1X106cells/ml) and three groups of rat were compared (control group, only LPS group, hMSC treatment in LPS group) through histopathological analysis after 12hr. Compared to only LPS-treated rats, hMSCs treatment in LPS-treated rats significantly showed increase of EBA-immunoreactive cells and reduction of Evans blue- immunoreactive cells in the substantia nigra. Interestingly, Compared to only LPS-treated rats, hMSCs treatment in LPS-treated rats significantly showed increase of the densities of astrocytes, assumed to significantly influence neurovascular structure and integrity and reduction of p-gp-immunoreactive cells, the one BBB transporter. Consequently, hMSC treatment significantly showed reduction of MPO-immunoreactive cells and increase of TH-immunoreactive cells in the substantia nigra. This study demonstrates that hMSCs treatment had a protective effect on neuroinflammation plays in the substantia nigra induced by LPS through inhibition of BBB permeability
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ABSTRACT ⅰ
TABLE OF CONTENTS ⅳ
LIST OF FIGURES ⅶ
Ⅰ. INTRODUCTION 1
A. Human mesenchymal stem cells exerts neuroprotection in an animal model
of double lesion-induced multiple system atrophy-parkinsonism (MSA-P) 2
B. Inhibition of blood brain barrier (BBB) permeability by hMSCs 3
Ⅱ. MATERIALS AND METHODS 5
A. MATERIALS 5
1. Antibodies 5
B. METHODS 5
1. Animal studies 6
2. Isolation of hMSC 6
3. Behavioral test 7
4. Extravasation of Evans Blue Dye 7
5. Tissue preparation 8
6. Immunohistochemistry 8
7. Western blot analysis 9
8. Stereological cell counts 10
9. Statistical analysis 11
Ⅲ. RESULTS 12
A. Human mesenchymal stem cells exerts neuroprotection in an animal model of
double lesion-induced multiple system atrophy-parkinsonism (MSA-P) 12
1. Characterization of hMSCs 12
2. Recovery of motor behavior by hMSCs 12
3. Detection of hMSCs in the double toxin-treated SN and striatum 13
4. Histological analysis of transplanted hMSCs in double lesion-induced
MSA-P model 13
5. Effect of cell therapy with hMSCs on modulation of inflammation and gliosis
in animals treated with double toxins 14
6. Effect of cell therapy with hMSCs on modulation of cell death signaling pathway
15
B. Inhibition of blood brain barrier (BBB) permeability by hMSCs 15
1. Effect of hMSCs on loss of dopaminergic neuron and modulation of
inflammation in LPS-induced animal models 15
2. Effect of hMSCs on BBB permeability in LPS-induced animal models 16
3. Effect of hMSCs on modulation of the densities of astrocytes
at the SN of animals treated with LPS 16
4. Effect of hMSCs on modulation of P-glycoprotein at the BBB
of LPS-induced animal models 17
5. Effect of hMSCs on MPO-1 neutrophil infiltration in the SNpc 17
Ⅳ. DISCUSSION 33
A. Human mesenchymal stem cells exerts neuroprotection in an animal model of
double lesion-induced multiple system atrophy-parkinsonism (MSA-P) 33
B. Inhibition of blood brain barrier (BBB) permeability by hMSCs 36
Ⅴ. CONCLUSION 40
A. Human mesenchymal stem cells exerts neuroprotection in an animal model of
double lesion-induced multiple system atrophy-parkinsonism (MSA-P) 40
B. Inhibition of blood brain barrier (BBB) permeability by hMSCs 40
REFERENCE 41
국문요약 50
