摘要目的 观察不同浓度亚砷酸钠(NaAsO2)对人皮肤永生化角质形成细胞(HaCaT)角化相关基因和核转录因子红系相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)mRNA表达的影响.方法 用0.00(对照)、3.13、6.25、12.50、25.00、50.00、75.00、100.00 μmol/L NaAsO2处理HaCaT细胞48h,采用2-(2-甲氧基-4-硝基苯基)-3-(4-硝基苯基)-5-(2,4-二磺酸苯)-2H-四唑单钠盐(CCK-8)法检测细胞增殖率.根据细胞增殖率检测结果,选择0.00(对照)、6.25、12.50、25.00μmol/L NaAsO2处理HaCaT细胞48 h,采用实时荧光定量PCR法检测HaCaT细胞角蛋白1(Keratin1,K-1)、角蛋白10(Keratin10,K-10)、总苞蛋白(Involurin,Inv)、兜甲蛋白(Loricrin,Lor)和Nr2的mRNA表达.结果 与对照组(100.05％)比较,6.25、12.50、25.00、50.00、75.00、100.00 μmol/L NaAsO2组HaCaT细胞增殖率(83.06％、51.04％、39.52％、24.51％、16.99％、9.04％)明显降低,半数致死量为12.38 μmoL/L.与对照组(1.06±0.28、1.00±0.12、1.00±0.08)比较,6.25、12.50、25.00 μmol/L NaAsO2组HaCaT细胞K-1、Inv、Lor mRNA表达(0.08±0.04、0.13±0.12、0.05±0.03,0.47±0.11、0.21±0.09、0.10±0.15,0.50±0.27、0.31±0.10、0.57±0.23)明显降低(P均＜0.05),但K-10 mRNA表达呈现升高趋势,其中6.25μmol/L NaAsO2组(1.83±0.45)明显高于对照组(1.07±0.14,P＜ 0.05);而12.50、25.00 μmol/L NaAsO2组Nrf2 mRNA表达(0.13±0.07、0.69±0.33)明显低于对照组(1.00±0.09,P均＜0.05).结论 NaAsO2 可通过影响细胞角化相关基因和Nrf-2 mRNA表达,抑制HaCaT细胞增殖与角化.

abstractsObjective To observe the influences of different doses of sodium arsenite on mRNA transcription of keratinizing related and nuclear factor E2-related factor 2(Nrf2) genes in HaCaT cells.Methods Cell proliferation was evaluated by Cell Counting Kit-8(CCK-8) assay after the HaCaT cells were exposed to 0.00,3.13,6.25,12.50,25.00,50.00,75.00,100.00 μ mol/L sodium arsenite for 48 h,respectively.Based on the previous results of cell proliferation,0.00(control),6.25,12.50,and 25.00 μmol/L of sodium arsenite were selected to treat HaCaT cells for 48 h,respectively.The mRNA expression of keratin 1,keratin 10,involucrin,loricrin and Nrf2 were detected by real-time fluorescent quantitative PCR.Results Compared with the control group (100.05％),HaCaT cell proliferation rates(83.06％,51.04％,39.52％,24.51％,16.99％ and 9.04％) were significantly lower in 6.25,12.50,25.00,50.00,75.00 and 100.00 μ mol/L of sodium arsenite groups and the 50％ inhibiting concentration was 12.38 μmol/L.Compared with the control group( 1.06 ± 0.28,1.00 ± 0.12,1.00 ± 0.08),the mRNA expression of keratin 1,involucrin and loricrin (0.08 ± 0.04,0.13 ± 0.12,0.05 ± 0.03;0.47 ± 0.11,0.21 ± 0.09,0.10 ± 0.15; 0.50 ± 0.27,0.31 ± 0.10,0.57 ± 0.23) were significantly decreased(all P ＜ 0.05) in HaCaT cells treated with 6.25,12.50,25.00 μmol/L sodium arsenite,respectively.But keratin 10 mRNA expression showed a rise trend and the 6.25 μmoL/L sodium arsenite group (1.83 ± 0.45) was significantly higher than that of the control( 1.07 ± 0.14,P ＜ 0.05 ).The Nrf2 mRNA expressions of HaCaT cells in 12.50,25.00 μmol/L sodium arsenite groups(0.13 ± 0.07,0.69 ± 0.33) were significantly lower than that of the control ( 1.00 ± 0.09,all P ＜ 0.05 ).Conclusions The cellular proliferation and keratinization are decreased when HaCaT cells are exposed to sodium arsenite,which may be regulated by lowering Nrf2 mRNA transcription.