중증 천식환자의 가래와 말초혈액에서 자가소화작용의 역할: 새로운 치료의 표적
Autophagy mechanisms in sputum and peripheral blood cells of patients with severe asthma: a new therapeutic target
- 주제(키워드) Autophagy; eosinophils; epithelial cells; severe asthma; treatment
- 발행기관 아주대학교
- 지도교수 박해심
- 발행년도 2016
- 학위수여년월 2016. 2
- 학위명 박사
- 학과 및 전공 일반대학원 의학과
- 실제URI http://www.dcollection.net/handler/ajou/000000021457
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Background: Autophagy and genetic predisposition have been suggested to potentially play roles in the development of asthma. However, little is known about the role of autophagy in the pathogenesis of severe asthma. Objective: We compared autophagy in the sputum granulocytes, peripheral blood cells (PBCs), and peripheral blood eosinophils (PBEs) between patients with severe asthma and those with non-severe asthma and investigated the functional effects of autophagy. Methods: We enrolled 36 patients with severe asthma, 14 with non-severe asthma, and 23 normal healthy controls in this study. Sputum granulocytes, PBCs, and PBEs were isolated from each subject. Autophagy was evaluated based on the expression of microtubule-associated protein light chain 3 (LC3) by western blot, confocal microscopy, transmission electron microscopy, and flow cytometry. IL-8 levels were measured by ELISA. To induce autophagy, HL-60 cells, SAECs, and A549 cells were treated with IL-5, IL-1β, and TNF-α. To inhibit autophagy, PI3K inhibitors (LY29400 and 3-methyladenine [3-MA]) and hydroxychloroquine (HCQ) were used. Knockdown of ATG5 and Beclin-1 was performed in A549 cells, and the therapeutic effects of dexamethasone were evaluated. Results: Higher autophagy levels were noted in sputum granulocytes, PBCs, and PBEs from patients with severe asthma than from patients with non-severe asthma and healthy controls (P<0.05 for all). IL-5 increased autophagy levels in both PBCs and PBEs (P<0.05). 3-MA attenuated the increased expression of LC3-II and eosinophil cationic protein in HL-60 cells induced by IL-5 (P=0.034 for both). Dexamethasone did not affect autophagy levels in PBEs. IL-1β increased LC3-II expression and IL-8 production (P<0.01) in SAECs, and this was attenuated by LY294002, 3-MA, HCQ, and knockdown of ATG5 and Beclin-1 (in A549 cells) (P<0.01). Conclusions and clinical relevance: Autophagy could play a role in the pathogenesis of severe asthma. Autophagy modulation may be a novel therapeutic target for conventional therapy-resistant severe asthma.
more목차
Ⅰ. INTRODUCTION 1
II. MATERIALS AND METHODS 4
A. MATERIALS 4
1. Study subjects 4
2. Antibodies and reagents 4
B. METHODS 5
1. Cell culture 5
2. Sputum induction and granulocyte isolation 5
3. Human PBC isolation and autophagy induction 6
III. RESULTS 12
IV. DISCUSSION 24
V. CONCLUSION 30
REFERENCES 31
국문요약 36
