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

作　　者：马瑶 白雪[1] 李煜[2] 陈柔锦 吴军[1] 

机构地区：[1]新疆医科大学公共卫生学院,新疆乌鲁木齐830011 [2]新疆乌鲁木齐市新市区疾病预防控制中心免疫规划科,新疆乌鲁木齐830000 [3]新疆医科大学第六附属医院教学科研科,新疆乌鲁木齐830002

出　　处：《环境与职业医学》2017年第11期999-1003,共5页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金(编号:81560513);新疆医科大学研究生科研创新基金(编号:CXCY2017013)

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、29.0μmol/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Keap1、Nrf2、ERK、NF-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ERK、NF-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Nrf2、ERK和NF-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。[ Objective ] To investigate the effects of mixture of arsenic on Keapl/Nrf2-ARE and MAPK/ERK signaling pathways and NF-~B gene expression in human immortalized keratinocytes （HaCaT cells） inhibited by Keapl. [ Methods ] HaCaT cells were cultured for 72 h and then divided into a control group （normal HaCaT ceils wihtout exposure to arsenic）, a negative control group （suppressed by Keapl without exposure to arsenic）, and three mixed arsenic exposure groups （suppressed by Keapl and exposed to arsenic at 2.9, 5.8, and 29.0 gmol/L, respectively）. Cell growth was measured by MTT method. The mRNA expression levels of Keapl, Nrf2, ERK, and NF-r^B 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 （？＇=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 Keapl 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 increased 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-EB 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-r^B 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 Keapl inhibition, arse

关 键 词：砷 Keap1抑制 HaCaT细胞 MAPK/ERK 核转录因子ΚB 

分 类 号：R994.6[医药卫生—毒理学]