铁过载氧化应激模型的构建、检测与分析

目的: 建立正确的铁过载氧化应激模型及检测手段。 方法: 通过显微镜镜检、MTT法、结晶紫染色等方法测定Fe-NTA对肝星状细胞系LX-2及肝细胞系Chang liver增殖状态的影响;利用紫外可见全波长扫描方法分析Fe与酚红、MTT试剂及显色产物、结晶紫之间的颜色反应。 结果: MTT试剂及显色产物与Fe之间存在颜色反应,不能用于检测铁过载氧化应激模型;结晶紫不与Fe发生颜色反应,结晶紫染色可作为铁过载氧化应激模型的检测手段;酚红与Fe之间存在一定的颜色反应,在铁过载氧化应激模型中应使用无酚红培养基。 结论: 构建铁过载氧化应激模型应使用无酚红培养基,并采用结晶紫染色方法检测细胞毒性。

Objective: To develop a correct model of oxidative stress induced by iron-overload and find out the right testing methods. Methods: Mesuring the cytotoxicity of Fe-NTA on LX-2 and Chang liver cells via microscope examination,the MTT assay and the crystal violet staining;analyzing the possible color reaction among phenol,Fe,the MTT reagents and the chromogenic products,and the crystal violet using the UV-vis full-length scanning. Results: MTT assay can not be used in the oxidative stress model induced by iron-overload for the color reaction existing between Fe and the MTT reagents and the chromogenic products,while crystal violet staining could be the right detecting mean in this model since there is no color reaction between Fe and the crystal violet.Then,as phenol can also react with Fe to generate colored products,medium without phenol should be used in the iron-overload model. Conclusions: In order to obtain the correct and valid experimental data,the Crystal Violet staining method and the phenol-free medium should be used in the oxidative stress model induced by iron-overload.