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目的研究适量硒对砷诱导的人脐静脉血管内皮细胞株(HUVEC-304细胞)诱导型一氧化氮合酶(iNOS)和内皮型一氧化氮合酶(eNOS)表达的影响。方法调整HUVEC-304细胞密度为1.0×105/ml,分别加入终浓度为0.05、0.1、0.5、1.5、3.0、6.0、12.0μmol/L三氧化二砷以及0.1μmol/L亚硒酸钠+0.05、0.1、0.5、1.5、3.0、6.0、12.0μmol/L三氧化二砷染毒48 h,并设立对照(正常培养液)组。检测培养液中iNOS和eNOS的活力,采用逆转录聚合酶连反应(RT-PCR)测定iNOS mRNA和eNOS mRNA的表达。结果随着三氧化二砷染毒浓度的升高,HUVEC-304细胞培养物中eNOS的活力呈逐渐下降的趋势,而iNOS的活力呈逐渐上升的趋势。与对照组比较,1.5～12.0μmol/L三氧化二砷染毒组HUVEC-304细胞培养物中iNOS mRNA的表达均较高,eNOS mRNA的表达均较低,差异均有统计学意义(P<0.05,P<0.01)。与相应浓度三氧化二砷单独染毒组比较,0.05、0.1、1.5μmol/L三氧化二砷+0.1μmol/L亚硒酸钠联合染毒组HUVEC-304细胞培养物中的iNOS的活力及其mRNA的表达均较低,0.05、0.5、1.5μmol/L三氧化二砷+0.1μmol/L亚硒酸钠联合染毒组eNOS的活力及其mRNA的表达均较高,差异均有统计学意义(P<0.05)。结论硒对砷诱导HUVEC-304细胞iNOS和eNOS的异常表达具有一定的抑制作用。
Objective To investigate the effect of appropriate amount of selenium on the expression of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in arsenic-induced human umbilical vein endothelial cell line (HUVEC-304). Methods The density of HUVEC-304 cells was adjusted to 1.0 × 10 5 / ml, and the cells were treated with 0.05, 0.1, 0.5, 1.5, 3.0, 6.0, 12.0 μmol / L arsenic trioxide and 0.1 μmol / L sodium selenite, 0.5,1.5,3.0,6.0,12.0μmol / L arsenic trioxide exposure 48 h, and the establishment of the control (normal broth) group. The activities of iNOS and eNOS in culture medium were detected. The expression of iNOS mRNA and eNOS mRNA were detected by reverse transcription polymerase chain reaction (RT-PCR). Results As the concentration of arsenic trioxide increased, the activity of eNOS in HUVEC-304 cells decreased gradually, while the activity of iNOS increased gradually. Compared with the control group, the expression of iNOS mRNA and the expression of eNOS mRNA in HUVEC-304 cells cultured in 1.5 ~ 12.0μmol / L arsenic trioxide group were all higher than those in control group (P <0.05, P <0.01). Compared with the corresponding concentration of arsenic trioxide alone exposure group, iNOS activity and mRNA expression in HUVEC-304 cells treated with 0.05, 0.1, 1.5μmol / L arsenic trioxide combined with 0.1μmol / L sodium selenite Low, 0.05,0.5,1.5μmol / L arsenic trioxide combined with 0.1μmol / L sodium selenite eNOS activity and mRNA expression were higher, the difference was statistically significant (P <0.05). Conclusion Selenium can inhibit the expression of iNOS and eNOS in arsenic-induced HUVEC-304 cells to a certain extent.