납에 의한 혈관내피세포의 사멸기전 및 산화환원조절단백-1의 보호효과

다국어 초록 (Multilingual Abstract)

Lead (Pb) is generally known as toxic metal ions in the cardiovascular cellular system. The effect of Pb on the endothelial cell death and its underlying mechanisms was investigated in the cultured human umbilical vein endothelial cells. To known the protective role of apurinic/apyrimidinic endonuclease1/redox factor-1 (named as Ref-1), Pb-induced cellular response was investigated in the Ref-1-overexpressed endothelial cells by using adenoviral Ref-1.
The exposure of Pb induced endothelial cell death in a dose-dependent manner at the range of 1 to 300μM and Pb-induced endothelial cell death was time-dependency (12 hr ~ 48 hr). Pb (30μM) increased superoxide and hydrogen peroxide production, and decreased catalase expression in the cultured endothelial cells. Interestingly, Ref-1 was increased about 250 % by the Pb exposure for 48 hr in the total lysate of endothelial cells. Pb exposure induced cytoplasmic translocation of Ref-1 in the enhanced green fluorescent protein-conjugated Ref-1-transfected endothelial cells.
Recombinant adenoviral Ref-1 was successfully over-expressed into the endothelial cells at the 200 multiplicity of infection. Forced over-expression of Ref-1 with adenoviral Ref-1 significantly inhibited Pb-induced endothelial cell death which was measured with morphological change and MTT assay. Overexpression of Ref-1 also abrogated Pb-induced superoxide and hydrogen peroxide production. Catalase expression was up-regulated by the overexpression of Ref-1 even Pb induced downregulation of catalase in the endothelial cells.
Taken together, the exposure of Pb induced endothelial cell death via the production of reactive oxygen species which might be linked with down-regulation of catalase in the endothelial cells. Overexpression of Ref-1 inhibited Pb-induced endothelial cell death via up-regulation of catalase, suggesting Ref-1 is a candidate for a target protein against Pb-induced cellular injury.

목차 (Table of Contents)

• Ⅰ. 서론 1
• 1. 납이 혈관에 미치는 영향 1
• 2. 납과 ROS와의 관계 3
• 3. APE1/Ref-1의 세포 내 기능 4
• Ⅱ. 연구방법 5
• 1. 세포주 및 시약 5
• 2. Adenoviral transfection 6
• 3. Diydroethidium 염색을 이용한 활성산소 생성 측정 6
• 4. H2DCFDA염색을 이용한 H2O2 생성측정 7
• 5. pEGFP/APE1/Ref -1벡터의 제조 7
• 6. MTT assay 8
• 7. 웨스턴 블럿 8
• 8. 자료처리 9
• Ⅲ. 연구결과 9
• Ⅳ. 논의 36
• Ⅴ. 결론 45
• 참고문헌 47
• ABSTRACT 61