소장자료

The protective effect of docosahexaenoic acid on human retinal microvascular endothelial cells exposed to oxidative stress

  • 송원경
  • Graduate School, Yonsei University
  • 2011
The protective effect of docosahexaenoic acid on human retinal microvascular endothelial cells exposed to oxidative stress
  • 자료유형
    학위논문
  • 서명/저자사항
    The protective effect of docosahexaenoic acid on human retinal microvascular endothelial cells exposed to oxidative stress=산화 스트레스에 노출된 인체 망막 미세혈관 내피세포에서 DHA의 보호효과/Won Kyung Song.
  • 발행사항
    Seoul : Graduate School, Yonsei University, 2011.
  • 개인저자
    송원경
  • 형태사항
    43장 : ill ; 26 cm.
  • 학위논문주기
    학위논문(박사) --Graduate School, Yonsei University :Dept. of Medicine,2011.2
  • 언어
    영어

소장사항

소장정보
번호 소장처 청구기호 도서상태 반납예정일 신청/예약
1 연세의학도서관/학위논문서가/교내공개(PDF) T 대출불가(별치) -

초록

Ischemic-driven ocular neovascularization is the most common cause of blindness in all age groups. Ischemia in retinopathy leads to several harmful events, including formation of reactive oxygen species (ROS). ROS play an important role in angiogenesis through various redox sensitive cascades and by upregulating VEGF expression, the key molecule in angiogenesis. Docosahexaenoic acid (DHA) is a major dietary ω-3 long-chain polyunsaturated fatty acid and a major structural lipid of sensory and vascular retina. In this study protective mechanisms of DHA in human retinal microvascular endothelial cells (HRMECs) exposed to oxidative stress were assessed.Primary cultures of HRMECs were stimulated by various concentrations of hydrogen peroxide (H2O2) to establish oxidative stress condition. Maximum HRMEC proliferation was observed at 20µM concentration. This condition was used for further experiments to show how physiologic concentration of DHA protects oxidative stress induced HRMEC changes. DHA (10µM) suppressed exogenous H2O2 stimulated proliferation of HRMECs. Anti-proliferative intracellular signaling mechanism of DHA involved PKC/MEK1,2/ERK cascade and coupled to the transcription of c-myc gene. DHA (10 µM) showed a potent scavenging ability of H2O2 stimulated intracellular ROS in HRMECs. NF-κB is a redox-sensitive transcription factor which activates inflammatory responses. DHA (10µM) inhibited H2O2 induced NF-κB activation (RT-PCR and western blot). DHA further suppressed inflammation by inhibition of H2O2 induced TNF-α, IL-1β, and IL-6 mRNA, and COX-2 and iNOS. DHA significantly decreased VEGF transcription and NADPH oxidase expression which is increased under oxidative stress condition. Finally H2O2 induced in vitro angiogenesis of HRMECs were significantly suppressed by DHA.Current treatment methods of retinal neovascularization such as laser photocoagulation, topical steroid, or intravitreal injection of anti-VEGF agent have side effects and work for limited period of time. We have documented that DHA protects HRMECs exposed to oxidative stress through various pathways. Therefore DHA consumption may have additive protective effects in retinal neovascular diseases such as diabetic retinopathy and retinopathy of prematurity

목차

Abstract
I. Introduction
II. Materials and Methods
1. HRMEC culture and exposure to oxidative stress
2. Docosahexaenoic acid (DHA) treatment
3. Cell proliferation assay
4. Immunoblot analysis
5. RT-PCR analysis
A. Isolation of total RNA
B. cDNA synthesis
C. PCR analysis
6. Intracelluar ROS measurement
7. Nitrite assay
8. Determination of MMP-2 and -9 activities
9. Immunocytochemistry
10. In vitro angiogenesis/Determination of capillary density
11. Statistical analysis
III. Results
1. Oxidative stress condition establishment
2. The effect of DHA on HRMEC viability
3. The effect of DHA in H2O2-induced intracelluar oxidation of HRMECs
4. The effect of DHA on H2O2-induced inflammation of HRMECs
5. The effect of DHA on H2O2-induced angiogenesis in HRMECs
IV. Discussion
V. Conclusion

References
Abstract (In Korean)