Aldosterone-inducible genes in mineralocorticoid receptor expressed human endothelial cell lines

To elucidate the aldosterone-regulated genes in vascular endothelial cells, we developed human endothelial cell line (EAhy926) stably expressing human mineralocorticoid receptor by retroviral system (MR-EAhy). Then gene expression profile of MR-EAhy stimulated with or without aldosterone was compared using DNA microarray analysis. MR-EAhy were serum-deprived for 24h with 0.2% dextran-coated charcoal-treated FBS, and then stimulated with or without aldosterone (10-9 M) for 6 hr. Then cells were harvested for RNA extraction.

为阐明血管内皮细胞中受醛固酮（aldosterone）调控的基因，我们通过逆转录病毒系统（retroviral system）构建了稳定表达人盐皮质激素受体（mineralocorticoid receptor）的人内皮细胞系EAhy926，并将其命名为MR-EAhy。随后采用DNA微阵列（DNA microarray）分析，对比经醛固酮处理与未处理的MR-EAhy细胞的基因表达谱。将MR-EAhy细胞置于含0.2%葡聚糖活性炭处理胎牛血清（Fetal Bovine Serum, FBS）的培养基中进行24小时血清饥饿处理，随后分别以10⁻⁹ M醛固酮刺激或不做刺激，持续培养6小时，最终收集细胞以提取RNA。