硒对氟诱导人脐静脉血管内皮细胞iNOS和eNOS表达的影响

Effect of Selenium on Expression of eNOS and iNOS Induced by Fluoride in ECV-304

目的研究一定浓度的硒对氟诱导人脐静脉血管内皮细胞(ECV-304)内皮型一氧化氮合酶(eNOS)和诱导型一氧化氮合酶(iNOS)表达的影响。方法体外培养人脐静脉血管内皮细胞株,加氟组培养液中分别加入400、1 000、2 000μmol/L的氟化钠;加氟加硒组培养液中分别加入400、1 000、2 000μmol/L的氟化钠,同时加入100 nmol/L的亚硒酸钠;对照组细胞在无血清培养液中培养;连续培养24 h,检测培养液中的iNOS、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。结果与对照组比较,400、1 000、2 000μmol/L加氟组iNOS活力和iNOS mRNA的表达增强(P<0.01);而400、1 000、2 000μmol/L加氟组eNOS的活力和1 000、2 000μmol/L加氟组eNOSmRNA的表达降低(P<0.05,P<0.01);与相应剂量的加氟组(400、1 000μmol/L)比较,加硒加氟组iNOS活力和iNOS mRNA的表达降低(P<0.05),而eNOS的活力和eNOS mRNA的表达升高(P<0.05);而加硒加氟组(氟剂量:2 000μmol/L、硒剂量:100 nmol/L)的iNOS与eNOS的表达与相应剂量的加氟组(2 000μmol/L)比较,差异无统计学意义(P>0.05)。结论硒对氟诱导人脐静脉血管内皮细胞iNOS和eNOS表达产生影响,但是对高剂量氟组作用不强。

Objective To know the effect of selenium on expression of nitric oxide synthase（eNOS） and inducible nitric oxide synthase （iNOS） induced by fluoride in ECV-304 （human umbilical vein endothelial cells）. Methods The cells, ECV-304, were divided into two groups and treated as follows, one group was treated with sodium fluoride at different doses （0, 400, 1 000 and 2 000 μmol/L）, the other was treated with sodium fluoride at different doses （0, 400, 1 000 and 2 000 μmol/L）and added sodium selenite （100 nmol/L） respectively. 24 h of treatment, The activity of eNOS and iNOS were measured by NO and NOS assay kits and the expression of eNOS mRNA and iNOS mRNA was analyzed by RT-PCR. Results The activity of eNOS and the expression of eNOS mRNA were down-regulated by fluoride with a dose dependent manner, while the activity of iNOS and the expression of iNOS mRNA were up-regulated. Selenium showed a significant antagonism to low level fluoride treated groups. Conclusion Selenium can antagonize the effect of fluoride on the expression of iNOS and eNOS in the vein endothelial cells when the fluoride exposure level is not too high.