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Photoprotective Activity of Isorhamnetin in UVB-Damaged Human HaCaT Keratinocytes

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Alternative Title
자외선 B에 의해 손상된 인간 HaCaT 각질형성세포에서 isorhamnetin의 광 보호 활성
Abstract
본 연구의 목적은 인간 HaCaT 각질형성세포에서 자외선(UV) B의 노출로 인한 세포손상에 대하여 특정 약용 식물에서 분리된 플라보노이드, isorhamnetin (3-methylquercetin)의 광 보호 특성을 밝히는 것이다. Isorhamnetin은 인간 HaCaT 각질형성세포에서 UVB로 생성된 세포 내 활성산소종의 소거능을 나타내었고 UVB로 유도되는 자멸소체(apoptotic body)의 형성, 핵 분절화(nuclear fragmentation) 및 아포토틱 sub-G1 DNA 파편을 확연하게 감소시킴으로써 세포자멸사를 억제하였다. 또한, Isorhamnetin은 UVB 노출에 의한 지질, 단백질 및 DNA와 같은 거대분자의 산화손상을 감소시켰다. 이 외에도 isorhamnetin은 UVB로 유도된 미토콘드리아 기능장애로부터 HaCaT 각질형성세포를 보호하였다. 이러한 결과는 isorhamnetin이 UVB로 매개되는 피부 질환에 대한 효율적인 제제로 개발될 수 있는 특수성을 가지고 있음을 암시한다.
The purpose of this study was to characterize the photoprotectiveproperties ofisorhamnetin(3-methylquercetin), an isolated flavonoidfrom certain medicinal plants, against cell injury and damage inhuman HaCaT keratinocytes resulting from ultraviolet (UV) B exposure. Isorhamnetin exhibited scavenging capacity of UVB-generated intracellular reactive oxygen species and repressed UVB-facilitated apoptosis in the human HaCaT keratinocyte, as evidenced by decreasing apoptotic body formation, nuclear fragmentation, and apoptotic sub-G1 DNAfraction. Furthermore, isorhamnetin attenuated the oxidative related macromolecular damage of lipids, proteins, and DNA in response to UVB radiation. In addition, isorhamnetin salved HaCaT cells from UVB light triggered mitochondrial dysfunction. Taken together, these findings impliedthat isorhamnetin has the peculiarity to be developed as an efficientagent against UVB-mediated skin disease.
Author(s)
한하
Issued Date
2015
Awarded Date
2016. 2
Type
Dissertation
URI
http://dcoll.jejunu.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000007487
Alternative Author(s)
Han, Xia
Department
대학원 의학과
Advisor
현진원
Table Of Contents
1. Introduction. 1
2. Materials and Methods. 3
2-1. Reagents. 3
2-2. Electron spin resonance (ESR) spectrometry for superoxide anion and hydroxyl radical detection.. 3
2-3.Cell culture.. 4
2-4. Cell viability assay. 4
2-5. Measurement of intracellular ROS.. 4
2-6. Nuclear staining with Hoechst 33342 dye. 5
2-7. Determination of mitochondrial membrane potential (Δψm). 6
2-8. Detection of sub-G1 hypodiploid cells 6
2-9. Terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick end labeling (TUNEL) assay... 7
2-10. Single-cell gel electrophoresis (comet assay) 7
2-11. Lipid peroxidation assay 8
2-12. Protein carbonyl formation 8
2-13. Statistical analysis.. 8
3. Results.. 9
3-1. Isorhamnetin exhibits capacity of free radicals scavenging in cell-free system and attenuates UVB-generated ROS in HaCaT keratinocytes... 9
3-2. Isorhamnetin protects HaCaT keratinocytes against UVB-induced apoptosis. 16
3-3. Isorhamnetin inhibits UVB-induced mitochondrial dysfunction.. 22
3-4.IsorhamnetinprotectsHaCaTkeratinocytes againstUVB-facilitatedmacromoleculardamage.. 24
4. Discussion. 28
5. References 30
6. Abstract in Korean 35
7. Acknowledgements. 36
Degree
Master
Publisher
제주대학교 대학원
Citation
한하. (2015). Photoprotective Activity of Isorhamnetin in UVB-Damaged Human HaCaT Keratinocytes
Appears in Collections:
General Graduate School > Medicine
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