马瑶, 白雪, 李煜, 陈柔锦, 吴军. 混合砷对Keap1抑制细胞Keap1/Nrf2-ARE和MAPK/ERK信号通路的影响[J]. 环境与职业医学, 2017, 34(11): 999-1003. DOI: 10.13213/j.cnki.jeom.2017.17401
引用本文: 马瑶, 白雪, 李煜, 陈柔锦, 吴军. 混合砷对Keap1抑制细胞Keap1/Nrf2-ARE和MAPK/ERK信号通路的影响[J]. 环境与职业医学, 2017, 34(11): 999-1003. DOI: 10.13213/j.cnki.jeom.2017.17401
MA Yao, BAI Xue, LI Yu, CHEN Rou-jin, WU Jun. Effects of mixed arsenic exposure on Keap1-inhibited Keap1/Nrf2-ARE and MAPK/ERK signaling pathways[J]. Journal of Environmental and Occupational Medicine, 2017, 34(11): 999-1003. DOI: 10.13213/j.cnki.jeom.2017.17401
Citation: MA Yao, BAI Xue, LI Yu, CHEN Rou-jin, WU Jun. Effects of mixed arsenic exposure on Keap1-inhibited Keap1/Nrf2-ARE and MAPK/ERK signaling pathways[J]. Journal of Environmental and Occupational Medicine, 2017, 34(11): 999-1003. DOI: 10.13213/j.cnki.jeom.2017.17401

混合砷对Keap1抑制细胞Keap1/Nrf2-ARE和MAPK/ERK信号通路的影响

Effects of mixed arsenic exposure on Keap1-inhibited Keap1/Nrf2-ARE and MAPK/ERK signaling pathways

  • 摘要: 目的 探究混合砷染毒对Keap1抑制的人永生化角质形成细胞(HaCaT细胞)Keap1/Nrf2-ARE和MAPK/ERK信号通路的影响及对NF-κB基因表达的影响。

    方法 细胞培养72 h,分为空白对照组(正常HaCaT细胞未染砷组)、阴性对照组(Keap1基因抑制且未染砷组)和3个Keap1基因抑制且混合砷染毒组,混合砷染毒浓度为2.9、5.8、29.0 μmol/L;采用MTT法测定细胞生长情况;采用实时荧光定量PCR法测定HaCaT细胞Keap1Nrf2ERKNF-κB的mRNA表达水平。

    结果 混合砷染毒组与阴性对照组的细胞活力相比,差异具有统计学意义(F=483.9,P<0.05)。中、高浓度砷染毒组细胞活性受抑制,差异有统计学意义(P<0.05)。混合砷染毒组与阴性对照组的Keap1 mRNA、Nrf2 mRNA相比,差异具有统计学意义(F=5.73,P=0.012;F=318.56,P<0.05)。Nrf2 mRNA表达呈低浓度时促进(P=0.038),中、高浓度砷染毒时抑制(P=0.014、P=0.016)。混合砷染毒组与阴性对照组的ERKNF-κB mRNA相比,差异具有统计学意义(F=39.88,P<0.05;F=2 619.41,P<0.05)。低砷浓度染毒促进ERK基因表达(P=0.020),高砷浓度染毒则抑制其表达(P=0.003),差异有统计学意义。NF-κB 基因表达与阴性对照组相比,低、中砷浓度时促进(P=0.030、P=0.032),高砷浓度时抑制(P=0.013),差异有统计学意义。

    结论 Keap1抑制状况下,砷对HaCaT细胞中Nrf2ERKNF-κB 基因表达呈低浓度时促进、高浓度时抑制的双向调节作用。

     

    Abstract: Objective To investigate the effects of mixture of arsenic on Keap1/Nrf2-ARE and MAPK/ERK signaling pathways and NF-κB gene expression in human immortalized keratinocytes (HaCaT cells) inhibited by Keap1.

    Methods HaCaT cells were cultured for 72 h and then divided into a control group (normal HaCaT cells wihtout exposure to arsenic), a negative control group (suppressed by Keap1 without exposure to arsenic), and three mixed arsenic exposure groups (suppressed by Keap1 and exposed to arsenic at 2.9, 5.8, and 29.0 μmol/L, respectively). Cell growth was measured by MTT method. The mRNA expression levels of Keap1, Nrf2, ERK, and NF-κB in HaCaT cells were determined by real-time fluorescence quantitative PCR.

    Results Compared with the negative control group, the cell activities of the mixed arsenic exposure groups were statistically different (F=483.9, P < 0.05). The cell activities of the medium dose and high dose arsenic exposure groups were inhibited, and the difference was statistically different (P < 0.05). The mRNA expression levels of Keap1 and Nrf2 were different between the mixed arsenic exposure groups and the negative control group (F=5.73, P=0.012; F=318.56, P < 0.05). The expression of Nrf2 mRNA was in creased at low dose (P=0.038), and inhibited at middle and high doses (P=0.014, P=0.016). The differences of ERK and NF-κB mRNA expression levels between the mixed arsenic exposure groups and the negative control group were statistically significant (F=39.88, P < 0.05; F=2 619.41, P < 0.05). Low-dose arsenic exposure promoted ERK gene expression (P=0.020), while high-dose arsenic exposure inhibited the expression (P=0.003), with significant differences. Compared with the negative control group, the expression of NF-κB gene was promoted by low-dose and middle-dose arsenic exposure (P=0.030, P=0.032), and inhibited by high-dose arsenic exposure (P=0.013), with significant differences.

    Conclusion Under Keap1 inhibition, arsenic mixture could promote the expression of Nrf2, ERK, and NF-κB genes at low dose, and inhibit expression at high dose.

     

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