황칠나무 추출물의 항산화 및 항염증 활성
Anti-oxidant and Anti-inflammatory Activities of Dendropanax morbifera Extracts
- 주제(키워드) 황칠나무 , 항산화 , 항염증
- 발행기관 아주대학교
- 지도교수 문은표
- 발행년도 2017
- 학위수여년월 2017. 8
- 학위명 석사
- 학과 및 전공 일반대학원 생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000025830
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
In this study, Dendropanax morbifera, a plant species native to Southeast Asia and Jeju Island in Korea, was examined for several biological activities. Four different fractions of Dendropanax morbifera extracts were used, and the extracts were named DPLS 1, DPLS 4, DPLS 5 and DPLW 4 according to solvent polarity and extraction parts of the plant. For anti-oxidant activity six different anti-oxidant tests were performed using trolox, an analog substance of vitamin E, as a standard material. The results revealed that DPLS 4 and DPLW 4 were the most effective anti-oxidant fractions. The overall anti-oxidant capacity of DPLS 1 was very low, but the hydroxyl radical scavenging activity and inhibition of lipid peroxidation were very high. The anti-inflammatory effects of the four extracts were examined by treating each extract to RAW264.7 cells simultaneously with the treatment of LPS to induce inflammatory responses. DPLS 1 showed the highest inhibitory activity against Nitric Oxide(NO) secretion, and the other three extracts also significantly inhibited NO secretion. The activity of blocking the NF-κB signaling pathway was further analyzed for DPLS 1. The results confirmed that the expression of iNOS and COX 2 was very effectively reduced in LPS-stimulated RAW264.7. DPLS 1 also reduced the translocation of NF-κB from the cytoplasm to the nucleus and inhibited the phosphorylation of IκB. In addition, the ability of the extracts to inhibit NLRP3 inflammasome formation was also investigated. First, the amount of intracellular ROS, one of the major causes of inflammasome formation, was analyzed. DPLS 1, a fraction that was the most effective in inhibiting the NF-κB signaling pathway, and DPLW 4, the most anti-oxidative fraction, were chosen for this analysis. DPLS 1 significantly reduced intracellular ROS, whereas DPLW 4 reduced extracellular ROS but did not decrease intracellular ROS. It is assumed that the polarity difference of the two extracts may be the reason for this result. Changes in the amount of NLRP3 protein following DPLS 1 treatment were also confirmed by western blot analysis, and as a result, DPLS 1 markedly decreased the amount of NLRP3 protein. Therefore, DPLS 4, DPLS 5 and DPLW 4, which had high anti-oxidative activities, are likely to be developed as a skin-whitening or anti-aging agent. DPLS 1 has the potential to be developed as a natural drug that can prevent diseases caused by excessive inflammation.
more목차
Ⅰ. Introduction 1
Ⅱ. Materials and Methods 3
1. Preparation of Dendropanax morbifera Extracts 3
2. Characterization of Dendropanax morbifera Extracts 3
2.1 Measurement of Phenolic Compound Contents 3
2.2. Estimation of Anti-oxidant Activities 4
2.2.1 DPPH Radical Scavenging Assay 4
2.2.2 ABTS Radical Scavenging Assay 4
2.2.3 Estimation of Reducing Power 5
2.2.4 Hydrogen Peroxide Scavenging Assay 6
2.2.5 Hydroxyl Radical Scavenging Assay 6
2.2.6 Inhibition of Lipid Peroxidation 7
3. Cell Culture 8
4. Estimation of Cytotoxicity 8
5. Measurement of Secreted Nitric Oxide 8
6. Detection of Intracellular ROS 9
7. Detection of Extracellular ROS 9
8. Western Blot Analysis 10
9. Statistical Analysis 11
Ⅲ. Results 13
1. Fractionation of Dendropanax morbifera Extracts 13
2. Characterization of Dendropanax morbifera Extracts 13
2.1 Total Phenolic Compound Contents in Dendropanax morbifera
Extracts 13
2.2 Anti-oxidative Activities of Dendropanax morbifera Extracts 15
2.2.1 DPPH Radical Scavenging Activity 15
2.2.2 ABTS Radical Scavenging Activity 15
2.2.3 Reducing Power 15
2.2.4 Hydrogen Peroxide Scavenging Activity 16
2.2.5 Hydroxyl Radical Scavenging Activity 16
2.2.6 Inhibition of Lipid Peroxidation 16
3. Cytotoxicity of Dendropanax morbifera Extracts 24
4. Inhibitory Effects on NF-κB Signaling Pathway 26
4.1 Measurement of Secreted Nitric Oxide 26
4.2 Detection of iNOS and COX 2 27
4.3 Detection of NF-κB Translocation 27
4.4 Detection of Phosphorylated IκB 28
4.5 Morphological Changes of RAW264.7 Cells 28
5. Inhibitory Effects on Inflammasome Formation 34
5.1 Measurement of Intracellular ROS 34
5.2 Measurement of Extracellular ROS 34
5.3 Detection of NLRP3 34
Ⅳ. Discussion 39
Ⅴ. References 44
Ⅵ. 국문요약 48
