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目的探讨5脂氧合酶激活蛋白在人脐静脉内皮细胞氧化损伤中的机制。方法体外分离、培养人脐静脉内皮细胞,分别用10、50、100和200mg/L氧化型低密度脂蛋白处理细胞24h,采用酶联免疫吸附法检测细胞上清液白三烯C4水平,MTT法检测细胞活力,以及实时荧光定量PCR和Western blot检测细胞5脂氧合酶激活蛋白mRNA及蛋白表达的变化。结果100mg/L和200mg/L氧化型低密度脂蛋白作用人脐静脉内皮细胞24h细胞活力显著下降(P<0.01),白三烯C4分泌显著增加(P<0.01),回归分析显示白三烯C4释放量与氧化型低密度脂蛋白浓度显著相关(r=0.953,P<0.01)。PCR和Western blot分析发现,5脂氧合酶激活蛋白mRNA表达和蛋白表达均显著增强(P<0.01和P<0.05)。结论氧化型低密度脂蛋白促进人脐静脉内皮细胞分泌白三烯C4,导致人脐静脉内皮细胞氧化损伤可能与5脂氧合酶激活蛋白异常表达有关。
Objective To investigate the mechanism of 5 lipoxygenase activating protein (HUVEC) oxidative damage in human umbilical vein endothelial cells. Methods Human umbilical vein endothelial cells were isolated and cultured in vitro. The cells were treated with 10, 50, 100 and 200 mg / L oxidized low density lipoprotein for 24 h respectively. The level of leukotriene C4 in the cell supernatant was detected by enzyme-linked immunosorbent assay (ELISA) Method was used to detect cell viability, and the changes of 5-LOX mRNA and protein expression were detected by real-time fluorescence quantitative PCR and Western blot. Results The cell viability of human umbilical vein endothelial cells treated with 100 mg / L and 200 mg / L oxidized low density lipoprotein decreased significantly (P <0.01) and the secretion of leukotriene C4 significantly increased (P <0.01). Regression analysis showed that leukotriene There was a significant correlation between C4 release and oxidized low density lipoprotein (r = 0.953, P <0.01). PCR and Western blot analysis showed that mRNA and protein expression of 5 lipoxygenase activator protein were significantly increased (P <0.01 and P <0.05). Conclusions Oxidized low density lipoprotein may promote the secretion of leukotriene C4 from human umbilical vein endothelial cells, which may be related to the abnormal expression of 5-LOX in oxidative damage of human umbilical vein endothelial cells.