Anti-inflammatory effects of lipid extracted from Hemitripterus villosus eggs on RAW264.7 macrophages
삼세기 알 유래 지질의 RAW264.7 대식세포에서의 항염증효과
- 주제(키워드) 도움말 Anti-inflammation , Lipids , RAW264.7 , Hemitripterus villosus , Fatty acids
- 발행기관 강릉원주대학교 일반대학원
- 지도교수 도움말 Park Woo Jung
- 발행년도 2024
- 학위수여년월 2024. 2
- 학위명 석사
- 학과 및 전공 도움말 일반대학원 해양식품공학과
- 세부분야 해당없음
- 실제URI http://www.dcollection.net/handler/kangnung/000000011772
- UCI I804:42001-000000011772
- 본문언어 영어
초록/요약 도움말
The sea raven (Hemitripterus villosus) is a marine fish widely distributed in the North Pacific Ocean. Total lipids were extracted from H. villosus eggs, and then they were subsequently separated into three lipids: neutral lipids, glycolipids, and phospholipids. The four fractionated lipids were recognized as abundant SFAs (17.49%–25.16% of total detected fatty acids), MUFAs (55.63%–22.30%), and PUFAs (19.21%–56.40%). In the present study, the anti-inflammatory activities of the total lipids and fractionated lipids of H. villosus eggs were investigated on LPS-stimulated macrophages. RAW264.7 cells were treated with each lipid (100–400 μg/mL) to evaluate the cytotoxicity, the production of NO, and the expression of immune-associated genes and signaling pathways. The results show that all H. villosus lipids dramatically reduced NO generation in LPS-stimulated RAW264.7 macrophages without toxicity. They also downregulated the transcription levels of COX-2, IL-1β, IL-6, and TNF-α expression in a concentration-dependent manner and also decreased the CD86 expression. Furthermore, H. villosus lipids inhibited the phosphorylation of NF-κB-p65, ERK1/2, JNK, and p38 via the NF-κB and MAPK signaling pathways. Therefore, the total lipids and fractionated lipids like neutral lipids, glycolipids, and phospholipids, which are isolated from H. villosus eggs have strong anti-inflammatory effect.
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 system 1
2. Immune response 3
3. Lipids from natural marine products 4
4. Sea Raven, Hemitripterus villosus 6
6. References 7
Chapter 2. The anti-inflammatory effects of total lipids isolated from Hemitripterus villosus eggs 19
Abstract 19
Introduction 20
1. Materials and methods 21
1.1 Preparation of H. villosus eggs 21
1.2 Lipid extraction 21
1.3 Analysis of the fatty acid profiles 22
1.4 Cell culture and sample treatment 23
1.5 Sample treatment 23
1.6 Cell proliferation and nitric oxide (NO) production assay 23
1.7 RNA extraction and cDNA synthesis 23
1.8 Real-time polymerase chain reaction (real-time PCR) analysis 24
1.9 Western blot assay 25
1.10 RAW264.7 cell production of surface molecules 26
1.11 Statistic analysis 26
2. Results and discussion 28
2.1 Gas chromatography analysis of total lipids from H. villosus eggs 28
2.2 Effect of total lipids isolated from H. villosus eggs on cytotoxicity in RAW264.7 cells 30
2.3 Effect of total lipids from H. villosus eggs on NO production 32
2.4 Effect of total lipids from H. villosus eggs on gene expression 34
2.5 Effect of total lipids extracted from H. villosus eggs on NF‑κB and MAPK activation 36
2.6 Effect of H. villosus eggs total lipids on the expression of cell surface molecules 38
3. Conclusions 40
4. References 41
Chapter 3. The anti-inflammatory effects of neutral lipids, glycolipids and phospholipids isolated from Hemitripterus villosus eggs 47
Abstract 47
Introduction 48
1. Materials and methods 49
1.1 Lipid extraction and fractionation 49
1.2 Analysis of fatty acid profiles of fractionated lipids 49
1.3 Determination of nitric oxide (NO) production and cell viability assay 49
1.4 Real-time PCR assay 50
1.5 Western blot assay 50
1.6 Flow cytometry analysis 50
1.7 Statistical analysis 51
2. Results and discussion 52
2.1 Gas chromatography analysis of fractionated lipids 52
2.2 Inhibitory effect of neutral lipids, glycolipids, and phospholipids from H. villosus eggs on cytotoxicity 54
2.3 Inhibitory effect of neutral lipids, glycolipids, and phospholipids from H. villosus eggs on NO production 55
2.4 Inhibitory effect of neutral lipids, glycolipids, and phospholipids from H. villosus eggs on gene expression 56
2.5 Inhibitory effect of neutral lipids, glycolipids, and phospholipids from H. villosus eggs on NF-κB and MAPK activation 60
2.6 Effect of neutral lipids, glycolipids, and phospholipids from H. villosus eggs on the expression of cell surface molecules 62
3. Conclusions 65
4. References 66
Chapter 4. Summary 71
