去甲肾上腺素对血管平滑肌细胞A7r5基因表达谱的影响

目的: 探讨去甲肾上腺素刺激后A7r5血管平滑肌细胞基因表达谱的改变。方法: 应用放射配体结合实验明确A7r5细胞上肾上腺素受体(AR)的表达。采用基因芯片技术观察去甲肾上腺素引起的血管平滑肌细胞基因表达谱的变化。用荧光实时定量PCR验证α_1A－AR,α_1B-AR的mRNA表达变化。结果:A7r5细胞中有α₁-AR和β-AR表达,其最大结合容量(Bmax)分别为(45.0±11.8)fmol／mg蛋白和(17.4±2.5)fmol／mg蛋白。去甲肾上腺素刺激细胞24 h后,有75个基因表达发生明显改变,其中涉及细胞结构、防御、代谢及信号转导等多方面的基因。荧光实时定量PCR结果显示去甲肾上腺素刺激细胞24 h后α_1A－AR和α_1B-AR的mRNA表达增加,与基因芯片的结果一致。结论:去甲肾上腺素激动AR引起A7r5血管平滑肌细胞多种不同功能的基因表达发生改变,提示在血管平滑肌中AR可能通过基因水平的调控而介导多种生物学功能。

DNA microarray profiling to identify norepinephrine-response genes in A7r5 aortic smooth muscle cells

AIM: To define the gene expression changes of vascular smooth muscle cells (VSMCs) in response to norepinephrine (NE). METHODS: The expression adrenergic receptors (AR) were determined by radioligand binding assay in A7r5 cells. Gene expression profiles were identified by cDNA microarray after A7r5 cells were treated with NE for 24 h, and mRNA expressions of α_1A-AR and α_1B-AR were confirmed by real-time PCR. RESULTS: α₁-AR and β-AR existed in A7r5 cells. Seventy-five genes with changed expression in response to NE were screened out. These genes are involved in cell structure, cell/organism defense, metabolism, signal transduction and so on. α_1A-, α_1B-AR mRNA expression identified by microarray and realtime quantitive PCR displayed similar patterns. CONCLUSIONS: Gene expression profile in response to NE was analyzed comprehensively with the microarray technique. NE induces many kinds of different function genes in A7r5 cells, which may provide a novel insight into the particular role of NE that modulates multiple aspects of biological function in VSMCs.