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目的观察叔丁基对苯二酚(tert-butylhydroquinone,tBHQ)对皮肤角质细胞核因子Nrf2(transcription factorNF-E2-related factor 2)及其调控的下游靶基因NAD(P)H醌氧化还原酶[NAD(P)H:quinone oxidoreductase 1,NQO1]和血红素单加氧酶-1(heme oxygenase 1,HO-1)蛋白表达的影响。方法向HaCaT细胞中加入终浓度为50μmol/L的tBHQ溶液分别培养0(对照)、2、6、12、24、36 h;或分别加入终浓度为0(对照)、25、50μmol/L的tBHQ溶液培养24 h。采用western blot法检测细胞Nrf2、NQO1和HO-1的蛋白表达。结果与对照组比较,50μmol/L的tBHQ染毒2～36 h时HaCaT细胞Nrf2蛋白表达水平,染毒12～36 h时HaCaT细胞NQO1蛋白表达水平以及染毒6～36 h时HaCaT细胞HO-1蛋白表达水平均升高,差异有统计学意义(P<0.05或P<0.01)。25μmol/L和50μmol/L tBHQ分别染毒24 h后HaCaT细胞的Nrf2、NQO1和HO-1的蛋白表达水平均高于对照组,差异有统计学意义(P<0.01)。与25μmol/L tBHQ染毒组比较,50μmol/L tBHQ染毒组Nrf2和HO-1蛋白表达水平均下降,NQO1蛋白表达水平升高,差异有统计学意义(P<0.01)。结论 tBHQ能够诱导皮肤角质细胞的核转录因子Nrf2及其调控的下游靶基因NQO1和HO-1的蛋白持续表达,从而对皮肤角质细胞产生保护作用。
Objective To observe the effects of tert-butylhydroquinone (tBHQ) on the transcription factor NF-E2-related factor 2 (NADF) and its downstream target genes NAD (P) H quinone oxidoreductase (P) H: quinone oxidoreductase 1, NQO1] and heme oxygenase 1 (HO-1) protein expression. Methods The cells were treated with tBHQ at a final concentration of 50 μmol / L for 0, 6, 12, 24 and 36 h, respectively. HaCaT cells were treated with 0, tBHQ solution for 24 h. Western blot was used to detect the protein expression of Nrf2, NQO1 and HO-1. Results Compared with the control group, the expression level of Nrf2 protein in HaCaT cells from 2 to 36 h after exposure to 50 μmol / L tBHQ, the expression of NQO1 in HaCaT cells from 12 to 36 h and the HO- 1 protein expression levels were increased, the difference was statistically significant (P <0.05 or P <0.01). The protein expression levels of Nrf2, NQO1 and HO-1 in HaCaT cells treated with 25μmol / L and 50μmol / L tBHQ for 24 h were significantly higher than those in control group (P <0.01). Compared with the 25μmol / L tBHQ group, the expression of Nrf2 and HO-1 protein decreased and the expression of NQO1 protein increased in 50μmol / L tBHQ group (P <0.01). Conclusion tBHQ can induce the sustained expression of Nrf2 and its downstream target genes NQO1 and HO-1 in skin keratinocytes, thereby protecting keratinocytes in the skin.