RAW264.7 대식세포에서 Oncorhynchus mykiss 알에서 추출한 지질의 항염증 활성
Anti-inflammatory activities of lipid extracted from Oncorhynchus mykiss eggs in RAW264.7 macrophages
- 주제(키워드) 도움말 면역 , 항염증 , 지질
- 발행기관 강릉원주대학교 웰니스바이오산업학과
- 지도교수 도움말 박우정
- 발행년도 2022
- 학위수여년월 2023. 2
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 웰니스바이오산업학과
- 세부분야 해당없음
- 실제URI http://www.dcollection.net/handler/kangnung/000000011325
- UCI I804:42001-000000011325
- 본문언어 한국어
초록/요약 도움말
Oncorhynchus mykiss (Rainbow trout) is the most widely farmed fish in the world and belongs to the salmon family. The lipids extracted from O. mykiss eggs contain the largest amounts of polyunsaturated fatty acids (PUFAs), about 51.92 % of the total fatty acids, which are predominantly EPA (7.77%) and DHA (33.66%). In this study, the anti-inflammatory effects of O. mykiss lipids in LPS-stimulated RAW264.7 cells were studied. O. mykiss lipids reduced LPS-induced NO production and mRNA expression of inflammatory mediators (iNOS and COX-2) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α), as well as cell surface CD86 expression. In addition, O. mykiss lipids inhibited the phosphorylation of JNK, ERK, p38, and NF-κB p65. These results revealed that O. mykiss lipids inhibited inflammatory responses by suppressing the MAPK and NF-κB signaling pathways. Additionally, PUFAs are precursors of eicosanoids, which are potent lipid mediators and play a role in inflammation and immune activation. The anti-inflammatory effects of O. mykiss lipids via the eicosanoid synthesis pathways were evaluated. LPS-induced COX-1 and 5-LOX expression was inhibited by O. mykiss lipids, while anti-inflammatory cytokines (TGF-β, IL-10, IL-11, BMP4, and LEFTY2) were increased. O. mykiss lipids inhibited LPS-induced production of thromboxane and prostaglandins in the serum, as well as expression of COXs, which is related to the COX pathway. In the LOX pathway, LTB4 production and 5-LOX expression levels were decreased. Furthermore, O. mykiss lipids reduced EET production and CYP4A11 expression in the cytochrome P450 pathway. These results confirmed that O. mykiss lipids activate the eicosanoid synthesis pathway. Therefore, these results provide insight into the anti-inflammatory properties of O. mykiss lipids on macrophages and suggest that O. mykiss lipids may be useful in preventing inflammation.
more목차 도움말
Table of contents I
List of Tables V
List of Figures VI
Acknowledgements VIII
Abstract IX
Chapter 1.
Literature Review 1
1. Immune responses 1
2. Lipid and lipid mediators 2
3. Oncorhynchus mykiss 4
4. References 5
Chapter 2.
Anti-inflammatory activities of Oncorhynchus mykiss lipids in LPS-induced RAW264.7 cells through NF-κB and MAPK signaling pathways 12
Abstract 12
1. Introduction 13
2. Materials and methods 14
2.1 Oncorhynchus mykiss samples 14
2.2 Lipid extraction 14
2.3 Fatty acid profile and gas chromatography (GC) analysis 14
2.4 Cell culture 15
2.5 Sample treatment 15
2.6 Cell proliferation assay 15
2.7 Determination of NO production 16
2.8 Quantitative real-time PCR (qRT-PCR) 16
2.9 Western blot analysis 17
2.10 Pathway inhibition analysis 18
2.10 Flow cytometry analysis 18
2.11 Statistical analysis 18
3. Results and discussion 19
3.1 Fatty acid profiles of O. mykiss egg lipids 19
3.2 Effects of O. mykiss lipids on cell viability 21
3.3 O. mykiss lipids inhibit LPS-induced NO production 21
3.4 O. mykiss lipids inhibit LPS-induced expression of inflammatory mediators and cytokines 24
3.5 O. mykiss lipids inhibit LPS-induced NF-κB and MAPK activation 26
3.6 Effects of O. mykiss lipids with specific inhibitors in LPS-inducted RAW264.7 cells 28
3.7 O. mykiss lipids inhibits LPS-induced surface molecule expression 30
4. Conclusions 33
5. References 34
Chapter 3.
Anti-inflammatory activities of Oncorhynchus mykiss lipids in LPS-induced RAW264.7 cells through eicosanoid synthesis pathway 40
Abstract 40
1. Introduction 41
2. Materials and methods 42
2.1 Isolation of O. mykiss lipids and treatments 42
2.2 Quantitative RT-PCR analysis (qRT-PCR) 42
2.3 Determination of eicosanoid levels 43
2.4 Western blot analysis 44
2.5 Statistical analysis 44
3. Results and discussion 45
3.1 O. mykiss lipids inhibit the expression of COX-1, 5-LOX, and anti-inflammatory cytokines 45
3.2 Anti-inflammatory effects of O. mykiss lipids on cyclooxygenase pathways 47
3.3 Anti-inflammatory effects of O. mykiss lipids on lipoxygenase pathways 50
3.5 Anti-inflammatory effects of O. mykiss lipids on cytochrome p450 pathways 53
4. Conclusions 56
5. References 57
Chapter 4.
Summary 60
