H2O2에 의한 clone-1-5c-4 결막세포주의 apoptosis에서 Catechin의 보호효과 = The protective effect of Catechin on apoptosis induced by H2O2 in clone 1-5c-4 Conjunctival Cell Lines

다국어 초록 (Multilingual Abstract)

A free radical causes a disfunction on the cell's anti-oxidation system. So it plays an very important role in the induction process of a cell's apoptosis. Hydrogen peroxide (H2O2) which is one of the reactive oxygen species has been seen to cause various diseases, various cellular disinfections, gene transformation and cell death. The green tea's extract, polyphenolic catechin has an effect on all sorts of physiologically active functions, anti-fungal, anti-inflammation and anti-allergic functions and it is known as a nature anti-oxidant agent. The purpose of this study was to identify the protective effect of EGCG on the cell death induced by oxygen free radical. We measured the removal ability of free radicals by DPPH free radical scavenging assay and evaluated the oxygen free radical's quantity in the cell by DCFH-DA assay. We measured cell viability by MTT assay, released LDH activity and analyzed DNA fragmentation to check up a distinctive feature in cell death. The mRNA expression in the cell were examined by RT-PCR. A processing of EGCG with each concentration after inducing cell death shows that a quantity of oxygen free radical in cell decreased as the concentration falls and the effect of scavenging DPPH free radical and cell survival rate increased as concentration rises. A ladder feature of DNA fragmentation decreased as a concentration falls. We examined mRNA expression related to cell death. The studies show that mRNA expression of bcl-2 and bcl-xL that control anti-apoptosis increased as a concentration was raised. bax mRNA expression that control pro-apoptosis decreased. Our results suggested that EGCG have an antioxidant effect in conjuctival cells induced cell death by H2O2. EGCG increased the expression of mRNA related to anti-apoptosis so that EGCG could be useful in the prevention of H2O2-induced conjunctival cells damaged through induction of the anti-apoptotic factor.

목차 (Table of Contents)

• List of Figureⅲ
• 국문 초록ⅴ
• 서론1
• 재료 및 방법4
• 1. 세포배양4
• 2. 시약 및 재료4
• 3. EGCG와 H2O2의 처리농도5
• 4. 세포생존율 분석6
• 1) MTT assay6
• 2) Lactic acid dehydrogenase (LDH) activity6
• 5. DNA fragmentation 분석7
• 6. DPPH free radical 소거능 측정8
• 7. Intracellular ROS (reactive oxygen species) 측정8
• 8. 역전사중합연쇄반응(RT-PCR)9
• 9. 자료의 통계정리10
• 결과11
• 1. 세포생존율 분석11
• 1) MTT assay11
• 2) Lactic acid dehydrogenase (LDH) activity12
• 2. DNA fragmentation 분석18
• 3. DPPH free radical 소거능 측정20
• 4. Intracellular ROS (reactive oxygen species) 측정22
• 5. 역전사중합연쇄반응(RT-PCR)25
• 고찰27
• 참고문헌30
• 영문 초록40