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目的探讨氧化低密度脂蛋白对培养血管内皮细胞表达基因Rictor的影响。方法分离并培养人脐静脉内皮细胞,用不同浓度(10、20、40和80 mg/L)氧化低密度脂蛋白作用24 h后,用RT-PCR及Western blot检测Rictor的表达情况,并检测转染Rictor后,蛋白激酶Akt及eNOS磷酸化的变化,用硝酸还原酶还原法测定NO释放量。结果氧化低密度脂蛋白显著降低Rictor的mRNA及蛋白的表达量(P<0.01),使Rictor-mTOR复合物形成减少。转染Rictor后,不仅Rictor表达增加而且使蛋白激酶Akt及eNOS磷酸化增加;内皮细胞NO释放量增多(P<0.05)。结论影响内皮细胞表达Rictor,是氧化低密度脂蛋白诱发血管内皮细胞功能不良的重要机制之一。
Objective To investigate the effect of oxidized low density lipoprotein on gene expression of Rictor in cultured vascular endothelial cells. Methods The human umbilical vein endothelial cells were isolated and cultured. The expression of Rictor was detected by RT-PCR and Western blot after treated with different concentrations (10, 20, 40 and 80 mg / L) of oxidized low density lipoprotein for 24 h, After transfection of Rictor, the phosphorylation changes of Akt and eNOS were detected by NO reduction method. Results Oxidized low-density lipoprotein (LDL) significantly decreased the mRNA and protein expression of Rictor (P <0.01) and decreased the formation of Rictor-mTOR complex. Transfection of Rictor not only increased the expression of Rictor but also increased the phosphorylation of Akt and eNOS, while NO release increased in endothelial cells (P <0.05). Conclusion The expression of Rictor in endothelial cells is one of the important mechanisms of oxidative low density lipoprotein-induced dysfunction of vascular endothelial cells.