- 學位論文
高尿酸對視網膜色素上皮細胞ARPE-19的生長及凋亡作用分析
The effects of hyperuricemia on growth and apoptosis of ARPE-19 retinal pigment epithelial cells
摘要
痛風與老年性黃斑部病變皆為代謝綜合症的一部分，而高尿酸血症可能會構成老年性黃斑部病變的風險之一，並且發病率皆會隨年齡增長而提高，然而其相關機制還尚未明確。本研究探討高尿酸對ARPE-19視網膜色素上皮細胞之影響，以不同尿酸濃度(0.3、0.5、0.7 x10-2M)加入視網膜色素上皮細胞培養後檢查其作用。以Trypan blue與CCK-8 assay檢測高濃度尿酸的影響，發現細胞數目與存活率皆降低，顯示高濃度的尿酸對細胞造成傷害。另外，DPPH試驗顯示尿酸在細胞外具有抗氧化的能力，以Sudan-Black-B染色可見高濃度尿酸會導致脂褐素生成，進一步以TUNEL染色證實細胞受到高濃度尿酸影響而凋亡。高濃度尿酸亦使VEGF表現量有顯著性上升，推測在體內可能會導致濕性老年性黃斑部病變，而Ang-Ⅱ的表現量未有變化。本研究初步證實高尿酸會損傷視網膜色素上皮細胞，但其中詳細病理發生機制及是否會在體內導致老年性黃斑部病變還需後續進一步探討。
並列摘要
Gout and age-related macular degeneration are both part of the metabolic syndrome, and hyperuricemia may constitute one of the risks of age-related macular degeneration, and the incidence of both increases with age. In this study, the effect of high uric acid on ARPE-19 retinal pigment epithelial cells was examined by adding different uric acid concentrations (0.3, 0.5, 0.7 x10-2M) to retinal pigment epithelial cells in culture. The cell numbers and survival rates were reduced in the results of the trypan blue and CCK-8 assays, indicating that high concentrations of uric acid caused cellular damages. In addition, DPPH assay showed that uric acid has antioxidant ability outside the cells, and Sudan-Black-B staining showed that high concentration of uric acid caused lipofuscin production. The high concentration of uric acid also caused a significant increase in VEGF expression, which may lead to wet age-related macular degeneration in vivo, while the expression of Ang-II did not change. This study primarily confirmed the detrimental effects caused by hyperuricemia. Nevertheless, whether high uric acid may cause in vivo age-related macular degeneration needs to be further investigated.