뇌 염증 반응에서 phosphatidylinositol 4-phosphat 5-kinases 의 조절 기능
Regulatory role of phosphatidylinositol 4-phosphate 5-kinases in brain inflammation
- 주제(키워드) PIP5KI , PI(4 , 5)P2 , astrocyte , Gmix , brain inflammaion
- 발행기관 아주대학교
- 지도교수 주일로, 이상윤
- 발행년도 2010
- 학위수여년월 2010. 8
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000010869
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2], mainly produced by the type I phosphatidylinositol 4-phosphate 5-kinase (PIP5KI) family members, PIP5KIα, PIP5KIβ and PIP5KIγ, is an important regulator of diverse cellular processes at the cell surface, such as cell survival, cell proliferation, phagocytosis, macropinocytosis membraned ruffles, ion channels transporters, cytoskeletal regulation and intracellular vesicle trafficking. Recently, it was reported that PI(4,5)P2 was important for neurodegenerative diseases. Nevertheless, expression and regulatory role of PIP5KIs in rat primary glia cell remain poorly understood. This present study has shown that PIP5KIα and PIP5KIγ are significantly upregulated in response to ganglioside mixture (Gmix) and lipopolysaccharide that are well known inflammatory stimulators. c-Jun N-terminal Kinase, phosphoinositide 3-kinase/Akt and extracellular signal-regulated kinase signaling pathways were responsible for the upregulation of PIP5KIα and PIP5KIγ. In addition, we have examined whether gene knockdown of PIP5KIα plays a role in the nuclear factor kappa B (NF-κB) signaling pathway. We found that PIP5KIα knockdown could inhibit the phosphorylation of NF-κB p65 in response to Gmix, leading to decrease in Gmix-induced proinflammatory cytokines, tumor necrosis factor α and interleukin-1β.These results suggest that PIP5KIα may act as a regulator of brain inflammation by modulating NF-κB signaling.
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ABSTRACT =ⅰ
LIST OF FIGURES = ⅲ
ABBREVIATION = ⅳ
Ⅰ. INTRODUCTION = 1
A. Phosphoinositide (PI) signaling networks = 1
B. Astrocyte & Microglia = 2
C. Cytokines = 2
D. Aims of study = 3
Ⅱ. MATERIAL AND METHOD = 5
A. Material = 5
1. Reagents = 5
2. Primary antibodies = 5
3. Construction of expressing plasmids = 6
B. Method = 6
1. Cell culture conditions and treatment = 6
2. Primary microglia cells and astrocyte = 6
3. Reverse transcription-Polymerase Chain Reaction (RT-PCR) = 7
4. Western blot analysis = 8
5. Transfection = 9
6. PI(4,5)P2 immunocytochemistry = 9
Ⅲ. RESULTS = 11
A. Upregulation of PIP5KIα and PIP5KIγ in astrocyte and microglia during inflammation response = 11
1. Induction of PIP5KIα and PIP5KIγ in rat primary astrocytes = 11
2. Induction of PIP5KIα and PIP5KIγ in rat primary microglia = 13
3. Induction of PIP5KIα enforced dose & time dependent manner during inflammation response = 15
4. Inflammation responses are accompanied by an increase in PI(4,5)P2 levels= 17
B. Induction mechanism of PIP5KIα and PIP5KIγ = 19
1. PI3K/AKT pathway & ERK signaling pathway = 19
2. c-Jun-N-terminal kinase (JNK) signaling pathway = 21
C. PIP5KIα is the mediator which is important for an inflammation response = 23
1. Transient transfection of the recombinant plasmids for gene silencing of PIP5KIα in rat astrocytes = 23
2. Loss of PIP5KIα gene inhibits proinflammatory cytokines induction = 25
3. Loss of PIP5KIαgene blocks IκB-α degradation and inhibits phosphorylation of NF-κB P65 = 25
Ⅳ. DISCUSSION = 30
Ⅴ. CONCLUSION = 33
REFERENCES = 34
국문요약 = 40
