3种抗氧化剂对亚砷酸钠染毒人膀胱上皮细胞血管内皮生长因子表达的拮抗作用研究

发布时间：2018-02-15 00:43

  本文关键词： 亚砷酸钠 膀胱上皮细胞 血管内皮生长因子 抗氧化　出处：《环境与健康杂志》2017年04期 　论文类型：期刊论文

【摘要】：目的探讨抗氧化剂褪黑素(melatonin,MEL)、N-乙酰半胱氨酸(N-acetyl cysteine,NAC)、维生素C(Vitamin,VC)对砷诱导人正常膀胱上皮(SV-HUC-1)细胞血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的拮抗作用。方法将处于对数生长期的SV-HUC-1细胞暴露于含终浓度分别为0(对照)、0.5、1、2、4、8、10μmol/L亚砷酸钠的F12K完全培养基染毒24 h,或者含终浓度分别为4、10μmol/L亚砷酸钠的F12K完全培养基染毒0(对照)、4、12、24、48、72 h;联合暴露组在加入含终浓度分为4μmol/L亚砷酸钠的F12K完全培养基前30 min分别加入1%二甲基亚砜(DMSO)和抗氧化剂MEL、VC、NAC(终浓度分别为0.5、1、1 mmol/L),染毒24 h。分别采用Western blot法和RT-PCR法检测SV-HUC-1细胞VEGF蛋白和mRNA的表达水平。结果 10μmol/L亚砷酸钠染毒组SV-HUC-1细胞VEGF mRNA的表达水平高于对照组,差异有统计学意义(P0.05);且随着亚砷酸钠染毒剂量的升高,SV-HUC-1细胞VEGF mRNA的表达水平呈上升趋势。与对照组比较,4μmol/L亚砷酸钠染毒12、24、48 h及10μmol/L亚砷酸钠染毒24和48 h后SV-HUC-1细胞VEGF mRNA的表达水平均升高,差异有统计学意义(P0.05);且随着亚砷酸钠染毒时间的延长,各剂量组SV-HUC-1细胞VEGF mRNA的表达水平均呈先上升后下降的趋势。与对照组比较,4μmol/L亚砷酸钠+DMSO染毒组SV-HUC-1细胞中VEGF蛋白的表达水平均增加,差异有统计学意义(P0.05)。与4μmol/L亚砷酸钠+DMSO染毒组比较,亚砷酸钠+NAC染毒组SV-HUC-1细胞中VEGF蛋白的表达水平较高,差异有统计学意义(P0.05);而亚砷酸钠与MEL和VC联合染毒组SV-HUC-1细胞中VEGF蛋白的表达水平无明显改变。结论砷能诱导人正常膀胱上皮细胞VEGF表达增加,NAC能增加VEGF表达。
[Abstract]:Objective to investigate the antagonistic effect of the antioxidant melatonin (melatonin), N-acetyl cysteine (N-acetyl cysteine) on the expression of vascular endothelial growth factor endothelial growth factor (VEGF) in human normal bladder epithelial cell line SV-HUC-1 induced by arsenic. Methods the SV-HUC-1 in logarithmic growth period was used to investigate the effect of melatonin on the expression of vascular endothelial growth factor (VEGF) in human normal bladder epithelial cell line SV-HUC-1. Methods the expression of vascular endothelial growth factor (VEGF) in human normal bladder epithelial cell line SV-HUC-1 was determined in logarithmic growth period. The cells were exposed to F12K complete medium containing 10 渭 mol/L sodium arsenite for 24 h or F12K containing final concentration of 4 10 渭 mol/L sodium arsenite for 0 h (control group 0 (control group 412124872 h); combined exposure group was exposed to F12K complete culture medium containing final concentration of 4 渭 mol/L sodium arsenite for 24 4872 h), and the combined exposure group was exposed to F12K complete medium containing 10 渭 mol/L sodium arsenite for 24 h. F12K complete medium with 4 渭 mol/L sodium arsenite was added with 1% dimethyl sulfoxide (DMSO) and antioxidant MELVCCONAC (0.5 渭 mol/L sodium arsenite) for 24 h. The expression of VEGF protein and mRNA in SV-HUC-1 cells were detected by Western blot method and RT-PCR assay, respectively, at the end concentration of 0.5 渭 mol / L ~ (-1) mmol-1 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1), respectively. Results the expression of VEGF mRNA in SV-HUC-1 cells exposed to 10 渭 mol/L sodium arsenite was higher than that in control group. The expression of VEGF mRNA in SV-HUC-1 cells increased with the increase of sodium arsenite dose. Compared with the control group, SV-HUC-1 cells were treated with 4 渭 mol/L sodium arsenite for 24 h and 10 渭 mol/L for 24 h and 48 h for 24 h and 48 h respectively. The expression level of VEGF mRNA was increased. The difference was statistically significant (P 0.05), and with the prolongation of sodium arsenite exposure time, The expression of VEGF mRNA in SV-HUC-1 cells increased at first and then decreased in each dose group. Compared with the control group, the expression of VEGF protein in SV-HUC-1 cells exposed to 4 渭 mol/L sodium arsenite DMSO was higher than that in control group. Compared with 4 渭 mol/L sodium arsenite DMSO group, the expression of VEGF protein was higher in SV-HUC-1 cells treated with sodium arsenite NAC than that in 4 渭 mol/L sodium arsenite DMSO group. The expression of VEGF protein in SV-HUC-1 cells exposed to sodium arsenite combined with MEL and VC did not change significantly. Conclusion arsenic can induce the expression of VEGF in human normal bladder epithelial cells and increase the expression of VEGF.
【作者单位】： 中国医科大学公共卫生学院地球化学性疾病研究室 "砷生物学作用评价及砷中毒防治"辽宁省重点实验室;
【基金】：国家科学自然基金(81072244)
【分类号】：R114
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本文编号：1512021