摘要
目的:本研究旨在探讨氧化应激在人表皮黑素细胞衰老中的作用。方法:以不同浓度的过氧化氢(0、200、400、600、800、1 000μmol/L)处理人黑素细胞5 d后,检测分析其细胞活力和凋亡。通过评估衰老相关指标β-半乳糖苷酶染色、细胞增殖、细胞周期分析、E-钙黏素以及衰老基因表达等改变,探讨低浓度过氧化氢(0~400μmol/L)对黑素细胞衰老进程的影响。结果:与对照组比较,当过氧化氢浓度为200μmol/L和400μmol/L时,细胞活力和凋亡未见显著性改变;高浓度的过氧化氢(600~1 000μmol/L)可以明显诱导黑素细胞凋亡;400μmol/L的过氧化氢则可使黑素细胞出现以细胞树突缩短、增殖减慢、β-半乳糖苷酶蓝染率增加、细胞周期阻滞于G2期、E-钙黏素表达降低及衰老相关蛋白p53和p21的表达增加等改变。结论:400μmol/L浓度的过氧化氢刺激能诱导黑素细胞提早衰老。
Objective：The aim of our research is to investigate the role of oxidative stress in cellular aging of human epidermal melanocytes. Methods：Cultured human melanocytes were subjected to different doses of H_2 _O2（0、200、400、600、800、1 000 μmol/L）for 5 d,after which cell viability and apoptosis were assessed.Next we investigated whether exposure to low levels of H2 O2（0-400 μmol/L）induces premature senescence via evaluating senescence-associated β-galactosidase staining,cell proliferation assay,cell cycle analysis,E-cadherin and senescent gene expression,etc. Results：Cell viability and apoptosis in melanocytes cultured with H_2O_2 at200-400 μmol/L showed no significant difference as compared with cells cultured without H_2O_2 and apoptosis of melanocytes progressively increased at high concentrations of H_2O_2（600-1 000 μmol/L）. Treatment with 400 u M H_2O_2 tended to exhibit dendrite retraction,increased SA-beta-gal activity,declined rate of proliferation,G2 cell cycle arrest,and changes in the expression levels of p53 and p21.We further show that pre-senescent melanocytes express markedly reduced levels of E-cadherin. Conclusion：Melanocytes undergo premature senescence under the stimulation of 400 μmol/L concentration of H_2O_2.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2018年第2期180-184,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金(81171517
81602774)
江苏省"333"工程项目(LGY2016009)
南京医科大学科技发展基金重点项目(2014NJMUZD044)
