PUKYONG

Chitosan oligosaccharides suppress oxidized LDL-induced foam cell formation and inflammatory response in RAW264.7 macrophages

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Alternative Title
RAW264.7 대식세포에서 키토산올리고당의 oxLDL의 의한 거품세포 형성 및 염증 반응 억제 효능
Abstract
Chitosan oligosaccharides (COS) are derived from chitosan or chitin which has been described to exhibit anti-inflammatory, antioxidant, anti-cancer properties and among others. The present of this research, COS had been used to investigate the inhibition of foam cell formation and anti-inflammatory effect in RAW264.7 macrophages stimulated with oxidized low-density lipoprotein (oxLDL). COS were found to have an inhibitory effect on foam cell formation. During ox-LDL stimulation of RAW264.7 macrophages, COS decreased cholesterol influx and SR-A1 and CD-36 protein expression levels while increasing cholesterol efflux and ABCA-1 and ABCG-1 protein expression levels. These findings imply that COS may have an inhibitory effect on foam cell formation. By reducing the production of reactive oxygen species (ROS), COS have shown promising anti-inflammatory properties by blocking the production of pro-inflammatory cytokines like tumor necrosis factor-alpha (TNF-α), and interleukin-1 (IL-1β), and interleukin-6 (IL6). Furthermore, COS in oxLDL-stimulated RAW264.7 macrophages increase heme oxygenase 1 (HO-1) expression whereas it suppresses iNOS and COX-2 expression. Moreover, COS significantly enhances the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2). According to these findings, COS may exert their protective effect by activating the Nrf2/HO-1 pathway. Key words: Chitosan oligosaccharides, foam cell formation, anti-inflammatory, oxLDL, RAW264.7 macrophages.
키토산 올리고당(COS)은 키토산 또는 키틴에서 파생되며 항염, 항산화, 항암 등의 특성을 나타내는 것으로 설명되었습니다. 본 연구의 현재, COS는 산화된 저밀도 지단백질(LDL)로 자극된 RAW264.7 세포에서 거품 세포 형성 억제 및 항염증 효과를 조사하는 데 사용되었습니다. COS는 거품 세포 형성에 억제 효과가 있는 것으로 밝혀졌습니다. RAW264.7 대식세포의 ox-LDL 자극 동안, COS는 콜레스테롤 유입과 SR-A1 및 CD-36 단백질 발현 수준을 감소시키면서 콜레스테롤 유출과 ABCA-1 및 ABCG-1 단백질 발현 수준을 증가시켰습니다. 그 외에도 COS 처리는 용량 의존적 방식으로 oxLDL 유도 RAW264.7 대식세포에서 핵인자-카파 B(NF-kB) 핵 전위를 억제합니다. 이러한 발견은 COS가 거품 세포 형성에 억제 효과를 가질 수 있음을 의미합니다. 활성산소종(ROS)의 생성을 줄임으로써 COS는 종양 괴사 인자 알파(TNF-α), 인터루킨-1(IL-1β), 인터루킨-6(IL6). 또한, oxLDL로 자극된 RAW264.7 세포의 COS는 헴-옥시게나제-1(HO-1) 발현을 증가시키는 반면 iNOS 및 COX-2 발현을 억제합니다. 또한, COS는 핵 인자 적혈구계 2 관련 인자 2(Nrf2)의 핵 전위를 상당히 향상시킵니다. 이러한 발견에 따르면 COS는 Nrf2/HO-1 경로를 활성화하여 보호 효과를 발휘할 수 있습니다.
핵심어: 키토산올리고당, 거품세포형성, 항염, oxLDL, RAW264.7 대식세포
Author(s)
LE THI MY PHUONG
Issued Date
2022
Awarded Date
2022. 8
Type
Dissertation
Keyword
Library
Publisher
부경대학교
URI
https://repository.pknu.ac.kr:8443/handle/2021.oak/32665
http://pknu.dcollection.net/common/orgView/200000643385
Affiliation
Pukyong National University, Graduate School
Department
대학원 식품생명과학과
Advisor
JeaYoungJe
Table Of Contents
1. Introduction 1
1.1. Chitosan oligosaccharides (COS) 1
1.2. Foam cell formation 5
2. Materials and methods 11
2.1 Materials 11
2.2 Cell culture 11
2.3 Cytotoxicity assay 11
2.4 Preparation of oxidized LDL and determination of thiobarbituric acid-reactive substances (TBARS) 12
2.5 Oil red O staining assay 12
2.6 Determination of intracellular cholesterol content 13
2.7 Cellular cholesterol influx experiment 13
2.8 Cellular cholesterol efflux experiment 13
2.9 Determination of intracellular ROS formation using DCFH-DA labeling 14
2.10 Determination of pro-inflammatory cytokines 14
2.11 Western blot analysis 15
2.12 Statistical analysis 15
3. Results 16
3.1 Effect of COS on cytotoxicity in RAW264.7 macrophages 16
3.2 Effect of COS on Oil red O staining assay 16
3.3 Effect of COS on total cholesterol, free cholesterol and cholesterol ester content 16
3.4 Effect of COS on cholesterol influx and efflux 20
3.5 Effect of COS on cholesterol regulatory protein: ABCG-1, ABCA-1, SA-A1, and CD-36 expression 20
3.6 Effect of COS on pro-inflammatory cytokine production 24
3.7 Effect of COS on ROS measurement 24
3.8 Effect of COS on HO-1, COX-2, and iNOS expression in oxLDL-stimulated RAW264.7 macrophages 27
3.9 Effect of COS on Nrf2 nuclear translocation 27
4. Discussion 31
5 Conclusion 38
6. Abstract (Korean) 39
7. Reference 41
Acknowledgement 47
Degree
Master
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대학원 > 식품생명과학과
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