探討抗氧化劑減輕氯化鎳引起的視網膜色素上皮細胞毒性

中文摘要老年性黃斑部病變的機轉非常複雜而且不易治療。臨床上分為乾性和濕性兩種類型，通常氧化壓力、發炎或血管新生為關鍵致病因。氯化鎳存在環境汙染物或合金成分中常出現的一種化合物，往往造成細胞氧化壓力、人體過敏或癌症轉移情形。因此本實驗主要目的是探討氯化鎳是否造成視網膜色素上皮細胞氧化壓力及發炎現象，進而使用抗氧化劑或保健營養品來減緩不良細胞反應。我們的結果顯示利用MTT分析發現氯化鎳造成視網膜色素上皮細胞毒性，細胞活性氧化物增加且細胞型態變細長且排列分散。利用PI stain及流式細胞分析儀分析得知氯化鎳造成細胞週期停滯在G2/M時期。以定量PCR方式觀察到HIF-1α 、HO-1、HK2及細胞自噬蛋白基因表現增加。而用ELISA發現氯化鎳造成視網膜色素上皮細胞分泌IL-8及VEGF。利用葉黃素、花青素(Cyanidin 3-O glucoside ，C3G)、 N-乙醯半胱胺酸及Metformin進一步探討，發現花青素(C3G)有抑制VEGF蛋白表現和IL-8的發炎。氯化鎳處理視網膜色素上皮細胞應該是老年性黃斑部病變很好的細胞模型應用。

AbstractThe mechanisms of age-related macular degeneration (AMD) are complex and difficult to treat. Clinically, it is divided into two types including dry and wet. Usually oxidative stress, inflammation or angiogenesis are the key causes. Nickel chloride is a compound often found in environmental pollutants or alloy components, often causing oxidative stress in cells, human allergies, or cancer metastasis. Therefore, the main purpose of this experiment is to explore whether nickel chloride causes oxidative stress and inflammation in retinal pigment epithelial cells, and then use antioxidants or health supplements to slow down adverse cell reactions. Our results show that using MTT analysis, nickel chloride can cause toxicity to retinal pigment epithelial cells, with increased cell reactive oxides species (ROS) and cell morphology becoming elongated and dispersed. Using PI stain and flow cytometry analysis, it was found that nickel chloride can cause cell cycle arrest in the G2/M phase. Increased expression of HIF-1α, HO-1, HK2 and autophagy protein genes was observed by quantitative PCR. Using ELISA, it was found that nickel chloride caused retinal pigment epithelial cells to secrete IL-8 and VEGF. Further exploration using lutein, anthocyanin (cyanidin 3-O glucoside, C3G), N-acetylcysteine (N-acetylcysteine) and Metformin found that anthocyanin (C3G) can inhibit the expression of VEGF protein and IL -8 inflammation. Nickel chloride treatment of retinal pigment epithelial cells should be a good cell model application for age-related macular degeneration.