LPS에 의한 중뇌 도파민성 신경 세포 사멸 동물 모델에서 Transient receptor potential vanilloid 1의 신경 세포 보호 작용
Transient receptor potential vanilloid 1 exhibits neuroprotective effects in LPS-induced dopaminergic neuronal death in the suabstantia nigra in vivo.
- 주제(키워드) TPRV1 , Parkinson's disease , microglia , inflammatio , infiltration , M1 M2
- 발행기관 아주대학교
- 지도교수 백은주, 진병관
- 발행년도 2011
- 학위수여년월 2011. 2
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000011584
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Transient receptor potential vanilloid 1 Exhibits Neuroprotective Effects in LPS-induced Dopaminergic Neuronal Death in the Substantia Nigra in vivo Parkinson？s disease (PD) is a motor disorder which is progressive degeneration of the nigrostriatal dopaminergic pathway. There is no cure for PD and besides the etiology of this disease was limited understood. Neuroinflammation have been proposed to play a role in the pathogenesis of PD. Recent evidence implicates the transient receptor potential vanilloid subtype 1 (TRPV1) leads to neuroprotective effects by anti-inflammtory action. The main purpose of this thesis is to investigate the functional effects of capsaicin (CAP), the TRPV1 agonist, in LPS-induced in vivo PD model. To do that, we investigated the LPS-induced inflammatory response including dopaminergic (DA) neuronal degeneration, microglia activation, inflammatory mediator expression and infiltration of peripheral immune cells in the SN. Additionally, we surveyed whether CAP would have ability to DA neuronal protection against LPS and how CAP regulates the inflamed condition. We also confirmed whether these effects of CAP were mediated by TRPV1. Immunostaining, RT-PCR and western blot analysis showed that CAP attenuates LPS-induced production of proinflammatory mediators such as inducible nitric oxide synthase (iNOS) and ROS while CAP increases Arginase 1 and CD206, M2 macrophage markers. In addition, CAP protected nigral DA neurons from LPS-induced neurotoxicity. This neuroprotective effect was reversed by capsazepine, a TRPV1 antagonist, indicative of TRPV1 involvement. However CAP could not block the infiltration of peripheral immune cells. Based on these results, we found that CAP exhibits the neuroprotective effects through regulating the immune cell phenotypes. The present results suggest that CAP may be beneficial for the treatment of neurodegenerative diseases, such as PD that is associated with neuroinflammation via TRPV1.
more목차
I. INTRODUCTION 1
A. Parkinson's disease 1
1. The characteristics of PD 1
2. Inflammation in PD 2
3. LPS model of PD 5
B. Immune cells in CNS 7
1. The characteristics of microglia 7
2. Infiltration of peripheral immune cells in CNS 9
3. Polarization of the immune cells 11
4. iNOS versus Arginase 1 13
C. Capsaicin and their receptor TRPV1 14
1. Capsaicin 14
2. Transient receptor potential vanilloid subtype 1 (TRPV1) 15
3. The neuroprotective effects of CAP and TRPV1 16
D. Aims of this study 17
II. MATERIALS AND METHODS 18
A. Materials and methods 18
1. Stereotaxic surgery 18
2. Pharmacological treatments 18
3. Tissue preparation and immunohistochemistry 19
4. Double-immunofluorescence Staining 21
5. Stereological cell counts of DA neurons 23
6. Counts of immuno-positive cells 23
7. Polymerase chain reaction (RTPCR) 23
8. Western blot analysis 24
9. In situ detection of O2- production 26
10. Cortical and mesencephalic microglia cultures 26
11. Determination of NO 27
12. Statistical analysis 27
III. RESULTS 30
A. The features of intranigral injection of LPS in the SN 30
1. LPS induces degeneration of DA neurons and activation of microglia in the SN 30
2. LPS induces activation of microglia in the SN 34
3. LPS induces proinflammatory cytokines, iNOS and COX-2 in CD11b-ip cells in the SN 36
4. LPS induces infiltration of various peripheral immune cells in the SN 39
B. The neuroprotective effects of CAP 42
