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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 establish a correct model of iron overload oxidative stress and detection means. Methods: The effects of Fe-NTA on the proliferation of LX-2 cells and Chang liver cell line were detected by microscopic examination, MTT assay and crystal violet staining. The effects of Fe and N-oxide on the proliferation of hepatic stellate cell line Changli were assayed by UV- Red, MTT reagent and color products, crystal violet color reaction between. Results: There was a color reaction between MTT reagent and chromogenic product and Fe, which could not be used to detect iron overload oxidative stress. Crystal violet did not color react with Fe. Crystal violet staining could be used as a test method for iron overload oxidative stress model There was a certain color reaction between phenol red and Fe, and phenol red-free medium should be used in the model of iron overload oxidative stress. Conclusion: To establish a model of iron overload oxidative stress, phenol red-free medium should be used, and cytotoxicity should be detected by crystal violet staining.