高浓度胰岛素对无血清培养的人脐静脉内皮细胞一氧化氮产生的影响

目的　观察无血清培养条件下 ,高浓度胰岛素对人脐静脉内皮细胞 (HU VECs)产生一氧化氮 (NO即 EDRF)功能的影响 .方法　第 2～ 4代的 HUVEC以普通 DMEM培养液培养 3d后 ,试验组换用含 14.35 ,2 8.70和 5 7.40 nmol· L- 1重组人胰岛素的无血清无酚红 DMEM培养液培养 ,对照组采用普通无血清无酚红 DMEM培养液培养 .每 2 4h换液 1次至72 h,换液时利用 Griess反应比色法检测细胞培养上清中 NO代谢产物— NO2 - 离子的 2 4h累积量 .结果　无血清培养条件下 2 8.70和 5 7.40 nmol· L- 1 胰岛素培养 48h可以使HU VEC培养上清 NO2 -离子的 2 4h累积量显著减少 .对照组 (4 7.0± 4.0 ) nmol,2 8.70 nmol·L- 1 胰岛素组 (4 0 .9± 3.4)nm ol;5 7.40 nm ol· L- 1胰岛素组 (38.1± 1.9) nmol,P<0 .0 5 .结论　高浓度胰岛素 (>2 8.70 nmol· L- 1 )长时间 (>48h)作用可以减少无血清培养的 HUVEC NO的产生 .由于NO是心血管系统重要的保护因子 ,高浓度胰岛素可能通过本途径直接促进心血管系统疾病的发生和发展 .

High concentration insulin decreases nitric oxide production in serum-free cultured human umbilical vein endothelium cells

AIM　To evaluate the effects of high concentration insulin on the nitric oxide(NO) production of serum-free cultured human umbilical vein endothelium cells(HUVECs). METHODS　After 3 days' culture in common DMEM medium, some HUVECs were cultured in serum-free phenol red-free DMEM containing 14.35, 28.70 or 57.40 nmol*L-1 insulin, others cultured in common serum-free phenol red-free DMEM medium only. Culture medium was renewed every 24 h till 72 h. The 24 h accumulated amount of NO2-(transformed from NO) in culture medium was detected by Griess Reaction. RESULTS　The 24h accumulated amount of NO2- in culture medium decreased significantly after 28.70 or 57.40 nmol*L-1 insulin cultured for 48 h；control group （47.0±4.0）nmol；28.70 nmol*L-1 group （40.9±3.4）nmol； 57.40 nmol*L-1 group （38.1±1.9）nmol, P<0.05. CONCLUSION　Under serum-free conditions, when insulin is cultured in high concentration (>28.70 nmol*L-1) for a relative long period (>48 h), the NO production in HUVECs may decrease, because NO is a protective factor in cardiovascular system; high concentration insulin may do harm to the cardiovascular system directly.