阿魏酸钠对ox-LDL诱导血管内皮细胞凋亡的影响

A study of interference of ox-LDL induced apoptosis of human umbilical vein endothelial cells by sodium ferulate in vitro

目的观察阿魏酸钠(SF)对氧化修饰的低密度脂蛋白(ox-LDL)诱导血管内皮细胞凋亡的保护作用。方法将体外培养的人脐静脉内皮细胞(HUVEC)分为5组,分别为对照组,ox-LDL(50mg/L)组,SF低浓度(5μmol/L)组,SF中浓度(10μmol/L)组及SF高浓度(20μmol/L)组。SF低、中、高浓度组在加入含相应浓度的SF培养1h后,加入终浓度为50mg/L的ox-LDL。采用MTT比色法确定SF的作用浓度。分别采用Hochest 33258细胞核荧光染色法(定性)和流式细胞仪(定量)检测SF对ox-LDL致内皮细胞凋亡的影响。结果一定浓度的SF能够抑制ox-LDL引起的HUVEC存活率降低,对HUVEC损伤具有拮抗作用,40μmol/L以下是SF拮抗细胞损伤的最佳浓度范围。Hochest33258染色后ox-LDL组细胞核显示较强蓝色荧光,可见核固缩、凋亡小体等典型的凋亡形态学特征;加入20μmol/L SF后,细胞核基本恢复其正常形态。流式细胞术结果显示,加入低、中、高浓度的SF后,其细胞凋亡率分别为25.21%±4.07%、12.45%±2.65%、10.85%±1.23%,与ox-LDL组(31.14%±4.29%)比较均明显降低(P<0.05),且存在量效关系。结论 SF具有拮抗ox-LDL诱导的HUVEC损伤,减少血管内皮细胞凋亡的作用。

Objective To investigate the effect of sodium ferulate（SF） on apoptosis of vascular endothelial cells induced by oxidized low density lipoprotein（ox-LDL） in vitro.Methods Human umbilical vein endothelial cells（HUVEC） cultured in vitro were assigned into five groups： the control group,Ox-LDL（50mg/L） group,SF（5μmol/L） group,SF（10μmol/L） group and SF（20μmol/L） group.SF in the said concentration was added to the latter 3 groups,respectively,and they were then cultured for one hour,followed by the addition of ox-LDL（50mg/L）.The effect of SF on HUVEC was assessed by MTT,and that on the ox-LDL induced HUVEC apoptosis was assessed with Hochest 33258 nucleus fluorescent staining and flow cytometer.Results A certain concentration of SF may increase the survival rate of HUVEC after the treatment with ox-LDL,thus it protects the HUVEC from injury.SF concentration below 40μmol/L was the optimal concentration in the protection of the cells from injury.With Hochest 33258 staining,the nucleus in ox-LDL group showed a dark blue fluorescence,with the appearance of typical morphological features of apoptosis,such as karyopyknosis and apoptotic body,etc.In SF（20μmol/L） group,the nuclei returned to its normal morphology.Flow cytometry revealed that the apoptosis ratio in low,medium and high concentration of SF groups（25.21%±4.07%,12.45%±2.65% and 10.85%±1.23%,respectively） was obviously lower than that in ox-LDL group（31.14%±4.29%,P0.05）,and a dose-dependent relation was observed.Conclusion Sodium ferulate may protect the ox-LDL induced HUVEC from injury and decrease apoptosis of vascular endothelial cells.