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Hyperoside protects V79-4 lung fibroblast cells against oxidative stress-induced apoptosis via inhibiting reactive oxygen species and c-Jun N-terminal kinase pathway

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Author(s)
Piao, Mei-Jing
Issued Date
2008
URI
http://dcoll.jejunu.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000004242
Abstract
This study is to elucidate the cytoprotective effect of hyperoside (quercetin-3-D-galactoside) against hydrogen peroxide (H₂O₂) induced apoptosis and its underlying mechanisms. Hyperoside was found to scavenge intracellular reactive oxygen species (ROS), and increase catalase activity. Thereby hyperoside prevented lipid peroxidation and DNA damage, showing inhibition of thiobarbituric acid reactive substance (TBARS) formation, inhibition of comet tail, decreased phospho-H2A.X expression, and inhibition of 8-hydroxy-2'-deoxyguanosine (8-OHdG) formation. Hyperoside protects cell death of Chinese hamster lung fibroblast (V79-4) cells via inhibition of apoptosis induced by H₂O₂ as shown by decreased apoptotic nuclear fragmentation, decreased sub-G1 cell population, decreased DNA fragmentation, inhibited mitochondria membrane potential (△Ψ) loss. The protective effects were also accompanied by up-regulation of bcl-2, down-regulation of bax, and the inactivation of caspase 9, and 3. Hyperoside abrogated the activation of c-Jun N-terminal kinase (JNK), an apoptosis related signal pathway, and its downstream transcription factor, activator protein-1 (AP-1). Taken together, these findings suggest that hyperoside protected H₂O₂ induced apoptosis of V79-4 cells by inhibiting ROS generation and JNK activation.
이 논문은 과산화수소(H₂O₂로 유도된 세포 사멸 및 그 메커니즘에 대한 hyperoside (quercetin-3?D-galactoside)의 세포 보호 작용을 해명하였다. Hyperoside는 세포 내 활성 산소 종 (ROS)을 소거하고 카탈라아제의 활성을 증가시킨다는 것을 발견하였다. Hyperoside는 thiobarbituric acid 반응 물질 (TBARS) 형성의 억제 작용, comet tail의 억제 작용, phospho-H2A.X 발현의 감소, 8-hydroxy-2'-deoxyguanosine (8-OHdG) 형성의 억제 작용을 나타냄으로써 지질 과산화 및 DNA 손상을 예방하였다. Hyperoside는 H₂O₂에 의하여 유도된 apoptosis의 억제 작용, 즉 apoptotic 핵 조각의 감소, sub-G1 세포 군의 감소, DNA 조각의 감소, 미토콘드리아 막 전위 (△Ψ) 손실의 억제를 통하여 중국 햄스터 폐 섬유아 (V79-4) 세포의 세포 사멸을 억제하였다. 이 보호 작용은 또 bcl-2의 up-regulation, bax의 dawn-regulation 및 caspase 9 또는 3의 비활성화 작용도 나타내었다. Hyperoside는 apoptosis와 관련되어 있는 신호 통로인 c-Jun N-terminal kinase (JNK)의 활성 및 그 하류 전사 인자, 활성 단백질-1(AP-1) 을 억제하였다. 상술한 바와 같이 이러한 연구 결과들로부터 hyperoside는 ROS의 생성 및 JNK 활성화의 억제를 통하여 H₂O₂로 유도된 V79-4 세포의 apoptosis를 억제하였음을 알 수 있었다.
Alternative Title
활성 산소종 및 c-Jun N-terminal 키나아제 통로의 억제를 통하여 산화적 스트레스로 유도되는 V79-4 햄스터 폐 섬유아세포의 세포사멸에 대한 Hyperoside의 보호작용
Affiliation
제주대학교 대학원
Department
대학원 의학과
Advisor
현진원
Awarded Date
2008. 2
Table Of Contents
Ⅰ. INTRODUCTION = 1
Ⅱ. MATERIALS AND METHODS = 3
1. Reagents = 3
2. Cell culture = 4
3. Intracellular reactive oxygen species (ROS) measurement = 4
4. Western blot = 5
5. Catalase activity = 5
6. Lipid peroxidation assay = 6
7. Comet assay = 6
8. Immunocytochemistry = 7
9. 8-Hydroxy-2′-deoxyguanosine (8-OHdG) assay = 7
10. Cell viability = 8
11. Nuclear staining with Hoechst 33342 = 8
12. Flow cytometry analysis = 8
13. DNA fragmentation = 9
14. Mitochondria membrane potential (Dy) analysis = 9
15. Preparation of nuclear extract and electrophoretic mobility shift assay = 9
16. Transient transfection and AP-1 luciferase assay = 10
17. Statistical analysis = 10
Ⅲ. RESULTS = 11
1. Effect of hyperoside on radical scavenging activity and catalase activity = 11
2. Effect of hyperoside on lipid peroxidation and cellular DNA damage induced by H₂O₂ = 15
3. Effect of hyperoside on apoptosis induced by H₂O₂ = 20
4. Effects of hyperoside on H₂O₂ induced MAP kinase and AP-1 activation = 25
Ⅳ. DISCUSSION = 28
Ⅴ. REFERENCES = 30
Ⅵ. ABSTRACT IN KOREAN = 35
Ⅶ. 감사의 글 = 36
Degree
Master
Publisher
제주대학교 대학원
Citation
Piao, Mei-Jing. (2008). Hyperoside protects V79-4 lung fibroblast cells against oxidative stress-induced apoptosis via inhibiting reactive oxygen species and c-Jun N-terminal kinase pathway
Type
Dissertation
Appears in Collections:
General Graduate School > Veterinary Medicine
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