硒对体外培养血管内皮细胞的影响

Effects of Selenium on Endothelial Cell Growth Cultured in vitro

目的研究不同浓度的硒对体外培养人脐静脉血管内皮细胞(HUVEC-304)的影响。方法体外培养人脐静脉血管内皮细胞株,在培养液中加入不同浓度的亚硒酸钠(50、100、500、1000、2000nmol/L),连续培养48h后,以瑞氏-吉姆萨染色观察细胞形态,以四唑氮蓝(MTT)比色法检测细胞活性,并检测培养液中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、诱导型一氧化氮合酶(iNOS)、内皮型一氧化氮合酶(eNOS)的活力,丙二醛(MDA)、NO的含量。结果硒浓度为1000、2000nmol/L时,细胞出现损伤样改变。硒浓度为100nmol/L时,细胞活性高于正常对照组(P<0.05);硒浓度为1000、2000nmol/L时,细胞活性降低。与对照组比较,硒浓度为50～500nmol/L时,培养液中的SOD、GSH-Px活力升高,差异有统计学意义(P<0.05);硒浓度达2000nmol/L时,SOD、GSH-Px活力明显降低(P<0.01)。与对照组比较,MDA在硒低浓度时无显著变化,当硒浓度升高至1000、2000nmol/L时显著增高(P<0.05,P<0.01)。低浓度的硒增强eNOS活力,降低iNOS活力,降低NO浓度;高浓度的硒降低eNOS活力,增强iNOS活力,升高NO浓度。结论低浓度的硒对脐静脉血管内皮细胞有促进增生作用,高浓度的硒有损伤作用,并随剂量的增高而加重。

Objective To research the growth of human umbilical vein endothelial cell（HUECV） cultured with different dose of selenium. Methods HUVEC were cultured with selenium in various concentration （50、100、500、1000、2000 nmol/L） for 48 h in vitro. The morphological changes of cells were observed, activity of cell was measured by MTT method, and at the same time the levels of SOD, GSH-Px, MDA, NO, iNOS, eNOS in the culture were determined. Results At the doses of 1 000-2 000 nmol/L, the cells were damaged and revealed related morphological change. Cell activity was significantly higher at the dose of 100 nmol/L compared with that of control （P〈0.05）, and decreased with the increased doses from 1 000 nmol/L to 2 000 nmol/L. The SOD and GSH-Px activity increased significantly at doses of 50-500 nmol/L, whereas decreased at 2000 nmol/L, compared with those of control（P〈0.01）. The MDA content showed no significantly variation at low dose group, but increased at doses of 1 000, 2 000 nmol/L significantly（P〈0.05, P〈0.01）. At the low concentrations of selenium group, the activity of eNOS increased, and the activity of iNOS decreased, and the content of NO decreased.At the high concentrations of selenium group, the activity of eNOS decreased, and the activity of iNOS increased, and the content of NO increased. Conclusion Low concentrations of selesium can increase cell activity, whereas high concentration of selenium damage HUVEC. The HUVEC can be damaged more seriously by higher concentration of selenium.