마우스에서 생체 시계 유전자 Per2의 조절 장애가 DSS로 유도된 대장염에 미치는 영향에 관한 연구
Dysregulated Circadian Clock Gene Per2 Exacerbates DSS-Induced Colitis by Enhanced Pro-Inflammatory Cytokine Production
- 주제(키워드) Circadian Clock , Per2 , DSS-induced colitis , proinflammatory cytokine , Th17 cell
- 발행기관 아주대학교
- 지도교수 장선영
- 발행년도 2018
- 학위수여년월 2018. 2
- 학위명 석사
- 학과 및 전공 일반대학원 약학과
- 실제URI http://www.dcollection.net/handler/ajou/000000027081
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Circadian rhythm regulates various aspect of physiological response in the single cell as well as behavior of an individual. Period 2 (Per2) gene is a member of circadian clock genes regulating mammalian circadian rhythm. Recent studies have reported that circadian rhythm disruption is associated with increased complication of inflammatory bowel disease. However, the underlying mechanism is unclear how circadian clock genes affect intestinal inflammation. In this study, we investigated whether unregulated circadian clock in absence of Per2 affects inflammatory response using chemically induced colitis. Per2 knockout (KO) mice were more susceptible to DSS-induced colitis than heterozygote littermates when the mice were monitored by change of body weights, disease activity score, and survival rates. Colon and mesenteric lymph nodes were analyzed for inflammatory cytokines, regulatory T cells, monocytes and neutrophils. Even though regulatory T cells were not impaired under steady state, regulatory T cells in MLN and colon, and inflammatory cytokines, especially TNF-alpha and IL-6, were up-regulated in colon of Per2KO mice under inflammatory condition. Per2KO mice showed decreased monocytes populations, but neutrophils were accumulated. The accumulation of neutrophils could be due to increased IL-17-producing CD4+ T cells in the lamina propria of Per2KO mice under steady state. These results suggested that unregulated circadian clock in absence of Per2 enhanced inflammatory tone against microbial stimuli in the colon.
more목차
I. INTRODUCTION 1
II. METHOD 3
A. Mice 3
B. DNA extraction and PCR for genotyping 3
C. DSS induced colitis model 3
D. Mononuclear cell isolation 4
E. Histology 4
F. Flow cytometry 5
G. Cytokine bead assay 5
H. Statistics 5
III. RESULTS
A. Clock-/- mice were not susceptible to DSS-induced colitis 7
B. Cry2-/- mice were not susceptible to DSS-induced colitis 10
C. Per2-/- mice were susceptible to DSS-induced colitis 13
D. Proinflammatory cytokines are up-regulated in Per2-/- mice under DSS-induced intestinal inflammation 16
E. Proinflammatory cytokines are up-regulated in Per2-/- mice under steady state condition 18
F. CD4+ Treg in Per2-/- mice is impaired under DSS-induced intestinal inflammatory condition 20
G. CD4+ Treg in Per2-/- mice is not impaired under steady state 22
H. Per2-/- mice had increased neutrophils in the colon under DSS-induced intestinal inflammatory condition 24
I. Per2-/- mice expressed enhanced IL-17A expressing Th17 cell in the colon under steady state 26
IV. DISCUSSION 28
V. CONCLUSION 31
REFERENCES 32
KOREAN ABSTRACT 36
