同型半胱氨酸对人脐静脉内皮细胞株ECV-304生长的抑制作用

The inhibitory effect of homocysteine on the growth of ECV-304 cells

目的 :观察同型半胱氨酸 (Hcy)对人脐静脉内皮细胞株ECV 30 4细胞生长的影响 ,探讨高同型半胱氨酸血症 (HHCY)致动脉粥样硬化 (As)的机制。方法 :将不同浓度Hcy与ECV 30 4细胞体外共培养不同时间 ,倒置显微镜下观察细胞形态 ,MTT法检测细胞生长抑制率 ,3 H TdR掺入法检测细胞DNA合成。结果 :显微镜见对照组细胞呈单层鹅卵石样紧密排列 ,而实验组随Hcy浓度的升高及作用时间的延长 ,细胞排列杂乱 ,聚集成团。低浓度Hcy对细胞生长的抑制作用不明显 ,5 .0、10 .0mmol/LHcy对细胞生长呈现抑制作用 ,且两组浓度作用 72h的生长抑制率高于作用 4 8h(P <0 .0 1)。 5 .0、10 .0mmol/LHcy作用 4 8h、72h ,3 H TdR掺入抑制率高于相应对照组和低浓度组 ,且作用 72h的掺入抑制率显著高于 4 8h(P <0 .0 1)。结论 :Hcy对VEC具有直接损伤作用 ,可以时间及剂量依赖方式抑制ECV 30 4细胞的生长及DNA的合成。

Objective:To investigate the effect of homocysteine(Hcy) on ECV-304 cells growth and to probe into a possible mechanism by which hyperhomocysteinemia (HHCY) results in atherosclrosis.Methods:ECV-304 cells were incubated with media containing a range of concentrations of Hcy for different time,changes in the morphology of cells were observed under microscope;thiazolyl blue tetrazolium bromide(MTT) assay was performed to examine the growth inhibitory rate of cells; 3H-TdR incorporation test was used to examine DNA synthesis of ECV-304 cells.Results:ECV-304 cells were identified by their typical cobblestone morphology when viewed by light microscope in control group,but the arrange of cells became disorderly,congregate with the increase of concentration of Hcy and the extension of time in test groups.No difference was found among 0.1,1.0,2.5 mmol/L Hcy groups and control group,but incubation with 5.0,10.0 mmol/L Hcy produced a significant inhibitory effect compared with control group (P<0.05-P<0.01).Treated with 5.0 and 10.0 mmol/L Hcy for 48 hour and 72 hour,the inhibitory rate at 72 hour was significantly higher than that at 48 hour(P<0.01).Incubation with 5.0,10.0 mmol/L Hcy for 48 hour,and 72 hour,the inhibitory rate of DNA synthesis was significantly increased than that in control group and low concentration groups(P<0.05-P<0.01);and the inhibitory rate of groups which were incubated with 5.0,and 10.0 mmol/L Hcy for 72 hour,was higher than that for 48 hour group(P<0.01).Conclusions:Hcy had a direct injurious effect on ECV-304 cells,and an inhibitory effect on the growth and DNA synthesis of ECV-304 cells in time- and concentration-dependent manner.