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星形胶质细胞维持神经元微环境,给予营养和代谢支持,并调节其对损伤的反应。鱼藤酮特异阻断线粒体复合物I,长期暴露于鱼藤酮可能增加患帕金森病的几率,并引起帕金森综合征。然而,星形胶质细胞在鱼藤酮所致多巴胺能神经元损伤过程中的作用尚无报道。本研究采用多巴胺能神经元细胞系MN9D 细胞模型,将经过或未经过星形胶质细胞条件培养基处理的MN9D 细胞暴露于不同浓度的鱼藤酮中,用计数法测生长曲线,MTT 法测细胞活性,DCFH 染色流式细胞仪测氧化应激水平,比色法测还原型谷胱甘肽含量。结果显示,MN9D 细胞在条件和普通培养基培养条件下生长曲线无明显差别;鱼藤酮浓度依赖性地降低细胞活性;不同浓度鱼藤酮作用24、48 h 后,经条件培养基处理的细胞其活性显著高于普通培养基培养的细胞;不同浓度的条件培养基都有保护作用,纯的条件培养基保护作用稍弱;预先24 h 条件培养基处理或同时给予鱼藤酮和条件培养基处理都有保护作用,鱼藤酮作用12 h 后再给予条件培养基则无保护作用;经条件培养基处理的细胞氧化应激水平降低;另外,条件培养基提高了细胞内还原型谷胱甘肽含量,缓解了鱼藤酮所致的谷胱甘肽耗竭。结果提示,星形胶质细胞可保护MN9D 细胞抵抗鱼藤酮所致的氧化应激,还原型谷胱甘肽可能参与了该保护过程。
Astrocytes maintain neuronal microenvironment, provide nutritional and metabolic support, and modulate their response to injury. Rotenone specifically blocks mitochondrial complex I, prolonged exposure to rotenone may increase the chances of having Parkinson’s disease and cause Parkinson’s syndrome. However, the role of astrocytes in rotenone-induced dopaminergic neuron injury has not been reported yet. In this study, MN9D cell model of dopaminergic neuronal cell line was used to study the growth curve of MN9D cells treated with or without astrocyte conditioned media in different concentrations of rotenone. The growth curve was measured by counting method. The cell viability was measured by MTT assay DCFH staining flow cytometry was used to measure the level of oxidative stress and the content of reduced glutathione was measured by colorimetry. The results showed that there was no significant difference in growth curve between MN9D cells and normal medium. Rotenone decreased the cell viability in a concentration-dependent manner. After treated with different concentrations of rotenone for 24 and 48 h, the activity of MN9D cells in conditioned medium was significantly higher The cells cultured in normal medium showed protective effects in different concentrations of conditioned medium, while the protective effect of pure conditioned medium was weaker. Pretreatment with 24 h conditioned medium or both rotenone and conditioned medium showed protective effect, After rotenone was added for 12 h, there was no protective effect on the conditioned medium. The oxidative stress level of the cells treated with conditioned medium decreased. In addition, the conditioned medium increased the content of intracellular reduced glutathione and relieved the rotenone induced Glutathione depletion. The results suggest that astrocytes protect MN9D cells against the oxidative stress induced by rotenone, and reduced glutathione may be involved in this protective process.