Evaluation the role of oxidative stress,mitochondrial membrane potential andantioxidants on manganese-induced cytotoxicityin neuronal cell line

• Main Authors: Pei-Fen Wu, 吳佩芬
• Other Authors: Chang-Yu Chen
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/37633089762011681769

碩士 === 輔英科技大學 === 醫事技術系碩士班 === 98 === Manganese (Mn) is a necessary trace element for human. It plays an important role in brain development, normal biochemical reaction and in the maintenance of cell function. Mn is also an environmental risk factor of health. Excessive Mn can cause serious nervous system diseases, mobility, mental and emotional abnormalities. In order to understand the mechanism of Mn-induced neurotoxicity, we use PC12 cells (pheochromocytoma) as cell mode, to investigate the effect of Mn on cytotoxicity, oxidative stress and mitochondrial membrane potential. PC12 cell lines were treated with different concentration manganese chloride (MnCl2) ( 0－1000 μM) and incubated for 24－72 hours. Cell viability were measured by MTT assay (methyl thiazolyl tetrazolium). The result shows that MnCl2 has can decrease the viability of PC12 cells, not only show time-effect but also dosage-effect. Analysis the ROS generation by DCF-dAc method. MnCl2 would increase the intracellular content of oxidants by concentration- and time-dependent manner. The effect of MnCl2 on mitochondrial membrane potential was determined by flow cytometer after Rhodamine 123 stain. After treatment with MnCl2 were significantly decreased the mitochondrial membrane potential in PC12 cells. Moreover, pretreatment with various antioxidants glutathione, N-acetylcysteine, superoxide dismutase, catalase, mannitol) inhibited the ROS generation and the decrease of mitochondrial membrane potential induced by MnCl2. Mn- induced PC12 cells death were significantly prevented by pretreatment with either glutathione / N-acetylcysteine , or catalase / superoxide dismutase, or mannitol. The mechanism of MnCl2-induced activation of PC12 cells death signaling pathway involved not only the excess generation of different types of intracellular oxidants, but also induced lose of mitochondrial membrane potential as well.