砷对人永生化角质形成细胞核转录因子红细胞系-2p45相关因子-2和Kelch样环氧氯丙烷相关蛋白-1基因表达的影响

发布时间：2018-03-03 22:13

本文选题：亚砷酸钠　切入点：人永生化角质形成细胞　出处：《环境与健康杂志》2015年10期 　论文类型：期刊论文

【摘要】：目的探讨不同剂量亚砷酸钠对人永生化角质形成细胞(human keratinocytes cell,Ha Ca T)Kelch样环氧氯丙烷相关蛋白-1(Keap1)和核转录因了红细胞系-2 p45相关因子-2(Nrf2)基因表达的影响。方法将Ha Ca T细胞分别暴露于终浓度为0(对照)、1.30、3.25、6.50μmol/L的亚砷酸钠培养基中培养24、48、72 h。采用流式细胞仪检测Ha Ca T细胞凋亡率,采用实时荧光定量(real-time quantitative PCR)检测Ha Ca T细胞中Nrf2、Keap1 m RNA的表达水平。结果与对照组相比,1.30μmol/L亚砷酸钠染毒24 h后Ha Ca T细胞的凋亡率降低,而3.25、6.50μmol/L亚砷酸钠染毒48、72 h后Ha Ca T细胞的凋亡率升高,差异均有统计学意义(P0.05);且随着亚砷酸钠染毒时间的延长和染毒剂量的升高,Ha Ca T细胞的凋亡率均呈上升趋势。与对照组相比,1.30μmol/L亚砷酸钠染毒24、48、72 h和3.25μmol/L亚砷酸钠染毒24 h后Ha Ca T细胞中Nrf2 m RNA的表达水平均增高,而3.25、6.50μmol/L亚砷酸钠染毒72 h后Ha Ca T细胞中Nrf2 m RNA的表达水平均下降,差异有统计学意义(P0.05);且随着亚砷酸钠染毒时间的延长和染毒剂量的升高,Ha Ca T细胞Nrf2 m RNA的表达水平均呈下降趋势。与对照组相比,1.30、3.25μmol/L亚砷酸钠染毒72 h和3.25、6.50μmol/L亚砷酸钠染毒48 h后Ha Ca T细胞中Keap1 m RNA的表达水平均增高,差异有统计学意义(P0.05)。随着亚砷酸钠染毒时间的延长,1.30μmol/L亚砷酸钠染毒组Ha Ca T细胞中Keap1 m RNA的表达水平呈波动性上升,3.25μmol/L亚砷酸钠染毒组Ha Ca T细胞中Keap1 m RNA的表达水平呈逐渐上升趋势,6.50μmol/L亚砷酸钠染毒组Ha Ca T细胞中Keap1 m RNA的表达水平呈先上升后下降;随着亚砷酸钠染毒剂量的升高,24、72 h时Ha Ca T细胞中Keap1 m RNA的表达水平呈下降趋势,而48 h时Ha Ca T细胞中Keap1 m RNA的表达水平呈上升趋势。结论亚砷酸钠可抑制Ha Ca T细胞的生长,诱导凋亡和角化的发生,其机制可能与Nrf2和Keap1基因的表达有关。
[Abstract]:Objective to investigate the effects of different doses of sodium arsenite on the gene expression of human immortalized keratinocytes, human keratinocytes cell Ha Ca Tntnka-like epichlorohydrin associated protein -1keap1) and nuclear transcription factor -2 p45 related factor -2nrf2. Methods Ha Ca T was used to induce the expression of Ha-Ca T gene in human immortalized keratinocytes. The cells were exposed to sodium arsenite culture medium with a final concentration of 0 (1.30 ~ 3.256.50 渭 mol/L) for 24 ~ 4872 h. The apoptosis rate of Ha Ca T cells was detected by flow cytometry. The expression of Nrf2keap1 m RNA in Ha Ca T cells was detected by real-time quantitative. Results compared with the control group, the apoptosis rate of Ha Ca T cells was decreased after exposure to 1.30 渭 mol/L sodium arsenite for 24 h. However, the apoptosis rate of Ha Ca T cells was increased after exposure to 3.25 ~ 6.50 渭 mol/L sodium arsenite for 48 ~ 72 h. The difference was statistically significant (P 0.05), and with the prolongation of sodium arsenite exposure time and the increase of dose of sodium arsenite, the apoptotic rate of Ha-Ca T cells showed an increasing trend. Compared with the control group, sodium arsenite was exposed to sodium arsenite at 1.30 渭 mol/L for 2448.72h and 3.25 渭 mol/L for sodium arsenite. The expression of Nrf2 m RNA in Ha Ca T cells increased 24 h after toxicity. However, the expression of Nrf2 m RNA in Ha Ca T cells decreased 72 h after exposure to 3.25 ~ 6.50 渭 mol/L sodium arsenite. The difference was statistically significant (P 0.05), and with the prolongation of sodium arsenite exposure time and the increase of dose, the expression of Nrf2 m RNA in Ha Ca T cells decreased. Compared with the control group, sodium arsenite was exposed to 1.30 ~ 3.25 渭 mol/L for 72 h and 3.25 渭 mol/L 6.50 渭 mol/L. The expression of Keap1 m RNA in Ha Ca T cells increased 48 h after exposure to sodium acid. With the prolongation of sodium arsenite exposure time, the expression of Keap1 m RNA in Ha Ca T cells was increased with the prolongation of sodium arsenite exposure to 1. 30 渭 mol/L. The expression of Keap1 m RNA in Ha Ca T cells was increased by 3.25 渭 mol/L sodium arsenite. The expression of Keap1 m RNA in Ha Ca T cells increased at first and then decreased in sodium arsenite exposed to 6.50 渭 mol/L. The expression of Keap1 m RNA in Ha Ca T cells decreased with the increase of sodium arsenite exposure for 2472 h. The expression of Keap1 m RNA in Ha Ca T cells increased at 48 h. Conclusion Sodium arsenite can inhibit the growth of Ha Ca T cells, induce apoptosis and keratogenesis, and its mechanism may be related to the expression of Nrf2 and Keap1 genes.
【作者单位】：新疆医科大学劳动卫生与环境卫生学教研室;
【基金】：国家自然科学基金(81560513;81260410) 教育部高等学校博士学科点基金(20126517110002;20126517120003)
【分类号】：R114

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