目的 观察糖化低密度脂蛋白(gly-LDL)对人血管内皮细胞生长及抗氧化相关因子的影响,探讨糖尿病患者血管内皮损伤与gly-LDL的相关性及可能机制。方法 用完全培养基及不同浓度(G1组0.06 mg/L,G2组0.12 mg/L,G3组0.24 mg/L)的gly-LDL处理人脐静脉血管内皮细胞(HUVECs)24 h、48 h及72 h,设对照组(Ctr),取细胞培养上清液检测SOD和MDA水平,同时用细胞划痕实验观察gly-LDL对内皮细胞迁移功能的影响,用实时半定量PCR(realtime quantitative PCR,RT-q PCR)方法测定Mn-SOD和e NOS的mRNA的表达情况。结果 与对照组相比不同浓度的gly-LDL均可抑制血管内皮细胞的迁移功能。较低或较高浓度的gly-LDL作用于人脐静脉内皮细胞时,随时间的增加SOD的活性及MDA的含量是升高的;中间浓度的gly-LDL作用于人脐静脉内皮细胞时,随时间的增加SOD活性是逐渐下降的,MDA含量是先增高后降低的。各组Mn-SOD的mRNA表达水平48 h和72 h均下调,e NOS的表达水平于24 h和48 h均下调,72 h时上调。结论 Gly-LDL与糖尿病患者血管并发症可能存在一定的相关性,但不能肯定其是恶化因素之一。
Objective To investigate the effect of glycated low density lipoprotein(gly-LDL) on the growth and antioxidant related factors in endothelial cells, and investigate the correlation of gly-LDL with diabetes vascular endothelial injury and the possible mechanism. Methods Human umbilical vein endothelial cells ( HUVECs ) were treated with different concentrations ( 0. 06 mg/L, 0. 12 mg/L, 0. 24 mg/L) of gly-LDL for 24 h, 48 h and 72 h. Superoxide dismutase(SOD) and malondialdehyde (MDA) levels in the cell culture supernatant were detected. The migrate function of endothelial ceils was assessed by scratch test. Mn-SOD and eNOS mRNA expressions were determined by real-time quantitative PCR (RT-qPCR). Results Gly-LDL at different concentrations inhibited the migrate function of endothelial ceils. When HUVCCs treated with 0. 06 mg/L and 0. 24 mg/L gly-LDL, the activity of SOD and MDA were increased with the time; while treated with 0. 12 mg/L gly-LDL, the SOD activity was decreases with the time and MDA was decrease after initial increase. Mn-SOD mRNA expression level were down-regulated after 48 h and 72 h in all gly-LDL treated groups. The expression of eNOS levels in 24 h and 48 h were down-regulated, but up-regulated in 72 h. Conclusion Glycated LDL can inhibit growth of endothelial cells and change the expression of antioxidant related factors, which may be associated with the pathogenesis of vascular complications in diabetic patients.