Thymelaea hirsuta (L.) Endl. extract inhibits the NLRP3 inflammasome by suppressing its ATPase activity
- 주제(키워드) NLPR3 인플라마좀 , 항염증 , IL-1β
- 주제(DDC) 570
- 발행기관 아주대학교 일반대학원
- 지도교수 Yong Hwan Park
- 발행년도 2026
- 학위수여년월 2026. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000036117
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
The NLRP3 inflammasome is a key component of the host defense system that orchestrates protective immune responses against a wide range of pathogens. At the same time, NLRP3 can be activated by disturbances in cellular homeostasis under non-infectious conditions, in which case it contributes to pathological inflammation. Consequently, the development of inhibitors that selectively modulate aberrant NLRP3 activation has emerged as a potential therapeutic strategy. In this study, we screened natural product–derived extracts for anti-inflammatory activity to identify NLRP3 inflammasome inhibitors with fewer adverse effects than synthetic therapeutics and simultaneously elucidated their mechanisms of action. Among the screened extracts, those derived from Smilax aspera L. and Thymelaea hirsuta (L.) Endl., collected in Tunisia, and Melampodium divaricatum (Rich.) DC., collected in Nicaragua, significantly reduced the secretion of the NLRP3 inflammasome– dependent pro-inflammatory cytokine interleukin-1β in macrophages. Their inhibitory effects were independent of upstream NF-κB signaling and potassium efflux and were not mediated by changes in intracellular reactive oxygen species (ROS) levels. Instead, these extracts suppressed the ATPase- dependent NLRP3 activation, thereby interfering with the assembly of inflammasome components. Furthermore, Thymelaea hirsuta (L.) Endl., which exhibited the strongest NLRP3-inhibitory activity in vitro, also reduced macrophage and neutrophil recruitment in a zebrafish inflammation model. Collectively, these findings provide a foundation for further mechanistic analysis of candidate anti- inflammatory compounds and suggest the potential of these natural extracts as effective and selective therapeutics for chronic inflammatory disorders. Keywords: NLRP3 inflammasome, Anti-inflammatory, IL-1β, Plant extract, NACHT domain, Smilax aspera, Thymelaea hirsuta, Melampodium divaricatum.
more목차
Ⅰ. Introduction 1
Ⅱ. Materials and Methods 3
1. Reagents and materials 3
2. In vitro experimental procedures 4
2.1 Cell culture and stimulation 4
2.2 Cell viability assay 4
2.3 NF-κB reporter gene assay 5
2.4 Cytoplasmic and nuclear protein fractionation 5
2.5 Intracellular ROS measurement 5
2.6 ASC oligomerization and speck immunofluorescence . 6
2.7 Measurement of NLRP3 ATPase activity 6
3. Statistical analysis 6
4. In vivo experimental procedures 7
4.1 Zebrafish husbandry and TMH preparations 7
4.2 Embryo survival experiments 7
4.3 Inflammation induction and TMH treatment 7
4.4 Sudan Black B staining 7
4.5 Whole-mount in situ hybridization 8
Ⅲ. Results 9
1. Screening of candidate compounds identifies SAP, TMH, and MelD as NLRP3 inhibitors 9
2. SAP, TMH, and MelD exhibit no cytotoxicity in macrophages 14
3. SAP, TMH, and MelD inhibit IL-1β release mediated by the NLRP3 inflammasome in mouse J774A.1 cells 16
4. SAP, TMH, and MelD inhibit IL-1β release mediated by the NLRP3 inflammasome in human THP-1 cells 18
5. SAP, TMH, and MelD selectively target the NLRP 3 inflammasome and do not inhibit AIM2 or NLRC 4 inflammasomes 20
6. SAP, TMH, and MelD do not affect NF-κB signaling 22
7. SAP, TMH, and MelD inhibit NLRP3 inflammasome activation independently of ROS and potassium efflux 24
8. SAP, TMH, and MelD inhibit ASC speck formation and ASC oligomerization 26
9. SAP, TMH, and MelD inhibit the ATPase activity of the NLRP3 28
10. TMH exhibits no toxicity in zebrafish embryos 30
11. TMH suppresses LPS-induced inflammation in zebrafish 32
Ⅳ. Discussion 35
Ⅴ. References 37
국문 초록 41
