同型半胱氨酸对ECV-304细胞NO和ROS形成的影响

目的: 观察同型半胱氨酸(Hcy)对ECV-304细胞产生一氧化氮(NO)及其对细胞内活性氧(ROS)产生的影响。方法: 将不同浓度的Hcy与ECV-304细胞体外共培养24h,硝酸还原酶法检测3种一氧化氮合酶(NOS)激动剂前后各组NO含量;流式细胞术检测细胞荧光强度,以反映ROS水平。结果: 0.50、1.00mmol/LHcy作用24h时NO量明显低于对照组(P<0.01);加入3种NOS激动剂刺激后,仍显示Hcy对ECV-304细胞释放NO反应的抑制作用,1.00mmol/LHcy组与对照组和低浓度组差异均有显著性(P<0.01)。0.50、1.00mmol/LHcy组的荧光强度明显高于对照组和低浓度组(P<0.01)。结论: Hcy可促进血管内皮细胞内ROS的产生,并抑制NO的形成,且呈剂量-效应关系。

The effect of homocysteine on the production of nitric oxide and reactivated oxygen species in ECV-304 cells

Objective:To observe the effect of homocysteine on the production of nitric oxide(NO) and reactivated oxygen species (ROS) in ECV-304 cells.Methods:ECV-304 cells were incubated with different of concentrations of Hcy for 24 hours.A nitric-acid reductase method was used to detect NO in different groups with and without three stimuli of nitric oxide synthase(NOS),and ROS in cells was detected by flow cytometry(FCM).Results:After cells were exposed to 0.5 mmol/L,1 mmol/L Hcy for 24 hours,NO in cultured media was significantly less than that in control group (P<0.01).Employing three stimuli for NOS,Hcy still resulted in a gradual decline in NO,and 1.0 mmol/L Hcy produced a statistical suppression of NO (P<0.01) compared with low concentration groups and control group.When incubated with 0.5 mmol/L,1.0 mmol/L Hcy fluorescence in cells detected by FCM was higher than that in control and low concentration groups (P<0.01).Conclusions:Hcy can dose-dependently restrain the excretion of NO and promote the production of ROS.