CAJ | 학술논문

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PIG124 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PIG124 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(P＜0.05)。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。
목적：건립불동농도적H2O2유도인흑소세포계PIG1산생양화응격상태적모형，위진일보연구백전풍양화응격발병궤제전정기출。방법분별이불동농도적H2O2처리인흑소세포계PIG124 h，광학현미경관찰흑소세포형태학변화，류식세포의검측세포조망정황급세포내활성양족(ROS)수평，CCK-8법검측처리후세포활성，류대류산파비타법검측세포지질과양화대사산물병이철(MDA)산생정황。결과수착작용농도적증가，흑소세포체적변소，수돌수량감소，장도축단，조망급배사축점증가；이0 mmol/L농도작위대조，0.50、0.75、1.00、1.25、1.50、2.00 mmol/L적H2O2분별처리인흑소세포계PIG124 h후，세포조망솔분별위6.8%、11.5%、15.6%、22.7%、38.7%화57.5%；세포활성수착작용농도적증가이축점강저，세포내ROS수평급MDA함량수착작용농도적증가이축점승고，1.00 mmol/L농도처리24 h후개시여대조조상비차이유통계학의의(P＜0.05)。결론해문모색출료H2O2인기인흑소세포양화응격손상적최가실험농도，병성공건립료인흑소세포양화응격손상모형。
ObjectiveTo establish an oxidative stress injury model of PIG1 cells with different concentrations of H2O2 and to further study the mechanism of oxidative stress in vitiligo.Methods After 24 hour treatment with different concentrations of H2O2, the morphological changes of PIG1 cells were observed by light microscopy. The intracellular ROS and apoptosis ratio of PIG1 cells were determined by lfow cytometry method. The cell viability was determined by CCK-8 assay. The malondialdehyde (MDA) content in PIG1 was determined by measuring thiobarbituric acid reactive substances to monitor lipid peroxidation.Results With the increasing of the concentration of H2O2, melanocytes turned smaller; the number of dendrites decreased and the length of them became shorter; cell death (apoptosis and necrosis) increased gradually. The apoptosis ratio of PIG1 cells induced by 0.50, 0.75, 1.00, 1.25, 1.50 and 2.00 mmol/L H2O2 were 6.8%, 11.5%, 15.6%, 22.7%, 38.7% and 57.5% respectively. Compared to the 0 mmol/L group, the viability of PIG1 cells was signiifcantly decreased when cells were treated by 1.0 mmol/L H2O2(P<0. 05). Compared with the control group, the intracellular ROS and the MDA content was signiifcantly elevated when cells were treated by 1.0 mmol/L H2O2(P<0.05). Conclusion The optimal concentration of H2O2 which could induce oxidative stress in PIG1 cells was determined and the cell injury model induced by H2O2 was successfully established.