过氧化氢诱导人黑素细胞氧化应激损伤模型的建立

发布时间：2018-09-07 13:47

【摘要】：目的建立不同浓度的H2O2诱导人黑素细胞系PIG1产生氧化应激状态的模型,为进一步研究白癜风氧化应激发病机制奠定基础。方法分别以不同浓度的H2O2处理人黑素细胞系PIG1 24 h,光学显微镜观察黑素细胞形态学变化,流式细胞仪检测细胞凋亡情况及细胞内活性氧簇(ROS)水平,CCK-8法检测处理后细胞活性,硫代硫酸巴比妥法检测细胞脂质过氧化代谢产物丙二醛(MDA)产生情况。结果随着作用浓度的增加,黑素细胞体积变小,树突数量减少,长度缩短,凋亡及坏死逐渐增加;以0 mmol/L浓度作为对照,0.50、0.75、1.00、1.25、1.50、2.00 mmol/L的H2O2分别处理人黑素细胞系PIG1 24 h后,细胞凋亡率分别为6.8%、11.5%、15.6%、22.7%、38.7%和57.5%;细胞活性随着作用浓度的增加而逐渐降低,细胞内ROS水平及MDA含量随着作用浓度的增加而逐渐升高,1.00 mmol/L浓度处理24 h后开始与对照组相比差异有统计学意义(P0.05)。结论该文摸索出了H2O2引起人黑素细胞氧化应激损伤的最佳实验浓度,并成功建立了人黑素细胞氧化应激损伤模型。
[Abstract]:Objective to establish a model of oxidative stress induced by different concentrations of H2O2 in human melanoma cell line PIG1, and to lay a foundation for further study on the pathogenesis of oxidative stress in vitiligo. Methods Human melanoma cell line PIG1 was treated with different concentrations of H2O2 for 24 h. The morphologic changes of melanocytes were observed by optical microscope. The apoptosis of melanocytes and the (ROS) level of intracellular reactive oxygen species were detected by flow cytometry and CCK-8 method respectively. The production of malondialdehyde (MDA) (MDA), a metabolite of lipid peroxidation, was detected by barbitol thiosulfate method. Results with the increase of action concentration, the volume of melanocytes became smaller, the number of dendrites decreased, the length of dendrites decreased, the apoptosis and necrosis increased gradually, and the H2O2 at 0 mmol/L concentration of 0 mmol/L was used to treat PIG1 cell line PIG1 for 24 h. The cell apoptosis rate was 6.8and 11.5g, respectively. The cell activity decreased gradually with the increase of the concentration of action, and the percentage of cell apoptosis was 38.7% and 57.5%, respectively. The level of ROS and the content of MDA increased gradually with the increase of action concentration. After 24 h treatment with 1.00 mmol/L concentration, there was a significant difference compared with the control group (P0.05). Conclusion the optimal concentration of oxidative stress on human melanocytes induced by H2O2 was found out, and the oxidative stress injury model of human melanocytes was successfully established.
【作者单位】：解放军第四军医大学西京皮肤医院;
【基金】：国家自然科学基金资助项目(81371739)
【分类号】：R-332;R758.41

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