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目的研究抗氧化剂对亚砷酸钠诱导的人膀胱上皮细胞系(SV-HUC-1)氧化应激相关通路的影响。方法取处于对数生长期的SV-HUC-1细胞,分别暴露于含终浓度为0(对照,F12K培养基)、4μmol/L亚砷酸钠及4μmol/L亚砷酸钠与丁硫氨酸亚砜亚胺(BSO,0.5 mmol/L)、褪黑素(0.5 mmol/L)或N-乙酰半胱氨酸(NAC,0.5 mmol/L)的培养基中,于培养16 h后,收集细胞进行转录因子NF-E2相关因子2(nuclear factor erythroid 2-related factor 2,NRF2)通路相关蛋白及其下游基因[血红素加氧酶1(heme oxygenase-1,HO1)、醌氧化还原酶(NAD(P)H-quinone oxidoreductase 1,NQO1)]蛋白表达水平的检测;于培养1 h后,收集细胞进行丝裂原活化蛋白激酶(mitogen-Activated Protein Kinase,MAPK)通路关键蛋白[p-细胞外信号调节激酶(p-ERK)、p-p38、p-c-Jun-N末端激酶(p-JNK)]表达水平的检测。结果与对照组比较,亚砷酸钠暴露SV-HUC-1细胞内NRF2、NQO1、HO1蛋白及p-ERK、p-p38、p-JNK蛋白的表达水平均较高;巯基耗竭剂BSO与亚砷酸钠联合暴露可加剧砷诱导的SV-HUC-1细胞内NRF2、NQO1、HO1和p-p38、p-JNK蛋白表达水平的升高,而抑制p-ERK的升高,差异均有统计学意义(P<0.05)。褪黑素与亚砷酸钠组及亚砷酸钠+NAC组SV-HUC-1细胞内NRF2蛋白的表达水平明显低于对照组和亚砷酸钠组;NQO1蛋白的表达水平仅明显高于对照组;HO1蛋白的表达水平明显高于对照组,而明显低于亚砷酸钠组,差异均有统计学意义(P<0.05)。褪黑素+亚砷酸钠组SV-HUC-1细胞内p-ERK、p-p38蛋白的表达水平明显低于对照组和亚砷酸钠组;p-JNK蛋白的表达水平明显高于对照组和亚砷酸钠组,差异均有统计学意义(P<0.05)。亚砷酸钠+NAC组SV-HUC-1细胞内p-p38蛋白的表达水平明显低于对照组和亚砷酸钠组;p-JNK蛋白的表达水平仅明显高于对照组;p-ERK蛋白的表达水平明显高于对照组,而明显低于砷暴露组,差异均有统计学意义(P<0.05)。结论抗氧化剂能够抑制亚砷酸钠急性暴露导致的SV-HUC-1细胞氧化应激相关通路的活化。
Objective To investigate the effects of antioxidants on the oxidative stress-related pathways of sodium arsenite-induced human bladder epithelial cell line (SV-HUC-1). Methods SV-HUC-1 cells in logarithmic growth phase were harvested and exposed to a concentration of 0 (control, F12K medium), 4μmol / L sodium arsenite and 4μmol / L sodium arsenite and butachlor ammonia After cultured for 16 h in medium of acid sulfoxide imine (BSO, 0.5 mmol / L), melatonin (0.5 mmol / L) or N-acetylcysteine The cells were harvested for the transcription factor NF-E2-related factor 2 (NRF2) pathway-related proteins and their downstream genes [heme oxygenase-1 (HO1), quinone oxidoreductase (NAD (P) H-quinone oxidoreductase 1, NQO1)] protein were detected; after 1 h of culture, the cells were harvested for mitogen-activated protein kinase (MAPK) pathway key protein [p- (P-ERK, p-p38, pc-Jun-N-terminal kinase (p-JNK)] expression levels were measured. Results Compared with the control group, the expressions of NRF2, NQO1, HO1 protein and p-ERK, p-p38, p-JNK protein in SV-HUC-1 cells exposed to sodium arsenite were all higher than those in control group Combination of sodium arsenite exposure increased the expression of NRF2, NQO1, HO1 and p-p38, p-JNK in arsenic-induced SV-HUC-1 cells and inhibited the increase of p-ERK Significance (P <0.05). The expression of NRF2 in melatonin and arsenite groups and in sodium arsenite + NAC group was significantly lower than that in control group and sodium arsenite group. The expression of NQO1 protein was only significantly higher than that in control group The expression of HO1 protein in the control group was significantly higher than that in the control group, but significantly lower than that in the sodium arsenite group (P <0.05). The expression of p-ERK and p-p38 protein in melatonin + sodium arsenite group was significantly lower than that in control group and sodium arsenite group; the expression level of p-JNK protein in SV-HUC-1 cells was significantly higher than that in control group Group and sodium arsenite group, the differences were statistically significant (P <0.05). The expression of p-p38 protein in SV-HUC-1 cells in sodium arsenite + NAC group was significantly lower than that in control group and sodium arsenite group; the expression of p-JNK protein was only significantly higher than that in control group; p-ERK The protein expression level was significantly higher than the control group, but significantly lower than the arsenic exposure group, the difference was statistically significant (P <0.05). Conclusion Antioxidants can inhibit the activation of oxidative stress-related pathways in SV-HUC-1 cells induced by acute exposure to sodium arsenite.