阿魏酸钠对ox-LDL诱导血管内皮细胞凋亡的影响

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。

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

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%,P<0.05),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.