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目的探讨亚砷酸钠对人角质形成(HaCat)细胞蛋白激酶B(PKB/Akt)及其下游信号因子IκB和NF-κB蛋白表达的影响。方法 HaCat细胞分别暴露于终浓度为0(对照)～100μmol/L的亚砷酸钠溶液6 h。采用Alamar Blue还原法检测细胞活力,采用Western Blot法检测细胞磷酸化Ak(tp-Akt)、磷酸化IκB(p-IκB)及胞核和胞浆中核转录因子NF-κB的蛋白表达水平。结果各浓度亚砷酸钠染毒组HaCat细胞内p-Akt、p-IκB蛋白以及胞核中NF-κB蛋白表达水平均显著升高,细胞活力和胞浆中NF-κB蛋白表达水平均显著下降,差异有统计学意义(P<0.05);且随着亚砷酸钠染毒浓度的升高,HaCat细胞内p-Akt、p-IκB蛋白以及胞核中NF-κB蛋白表达水平呈逐渐上升的趋势,细胞活力和胞浆中NF-κB蛋白表达水平呈逐渐下降的趋势。结论亚砷酸钠可使HaCat细胞活力明显下降;诱导HaCat细胞Akt蛋白的磷酸化活化,进而诱导其下游信号因子IκB磷酸化及核转录因子NF-κB入核表达。
Objective To investigate the effects of sodium arsenite on the expression of protein kinase B (PKB / Akt) and its downstream signal IκB and NF-κB proteins in human keratinocytes (HaCat). Methods HaCat cells were exposed to sodium arsenite solution at a final concentration of 0 (control) to 100 μmol / L for 6 h. Cell viability was measured by Alamar Blue reduction assay. The protein expression levels of phosphorylated Akt, phosphorylated IκB (p-IκB) and NF-κB in nucleus and cytoplasm were detected by Western Blot. Results The expressions of p-Akt, p-IκB protein and nuclear NF-κB protein in HaCat cells were significantly increased with different concentrations of sodium arsenite, and the cell viability and NF-κB protein expression in cytoplasm were significantly increased (P <0.05). The expression of p-Akt, p-IκB protein and nuclear NF-κB protein in HaCat cells gradually increased with the increase of sodium arsenite concentration Increasing trend, cell viability and cytoplasmic NF-κB protein expression showed a gradual downward trend. Conclusion Sodium arsenite can significantly reduce the viability of HaCat cells and induce the phosphorylation of Akt protein in HaCat cells, which in turn leads to the phosphorylation of IκB signaling and NF-κB nuclear translocation.