Candidate genes and mechanisms for 2-methoxyestradiol-mediated vasoprotection.
Candidate genes and mechanisms for 2-methoxyestradiol-mediated vasoprotection.
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DOI：
10.1161/hypertensionaha.110.152298
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发表时间：
2010-11
期刊：
影响因子：
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通讯作者：
Dubey RK
中科院分区：
文献类型：
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作者：
Barchiesi F;Lucchinetti E;Zaugg M;Ogunshola OO;Wright M;Meyer M;Rosselli M;Schaufelberger S;Gillespie DG;Jackson EK;Dubey RK
甲氧基乙烯（2-ME;雌二醇代谢物）抑制血管平滑肌细胞（VSMC）生长并保护动脉粥样硬化和血管损伤;然而，2-ME诱导这些作用的机制仍然不清楚。为了评估2-ME对调节VSMC生物学的生化途径的影响，我们采用高密度寡核苷酸微阵列来鉴定用2-ME急性（4小时）或长期（30小时）处理的培养的人女性主动脉VSMC中差异表达的基因。单基因分析和基因集富集分析（GSEA）均显示2-ME诱导的VSMCs中参与有丝分裂纺锤体组装和功能的基因下调。GSEA分析还鉴定了2-ME对调节细胞周期进程、细胞迁移/粘附、血管舒张、炎症和胆固醇代谢的基因的影响。转录变化与蛋白质表达的变化相关，包括细胞周期蛋白D1、细胞周期蛋白B1、cdk 6、cdk 4、微管蛋白聚合和胆固醇及类固醇合成的抑制以及环氧化酶-2（考克斯-2）和基质金属蛋白酶-1的上调。微阵列数据提示2-ME可激活VSMCs上的过氧化物酶体增殖物激活受体（PPARs），且2-ME与PPARγ激动剂罗格列酮具有相似的结构。然而，我们发现2-ME与PPARs的弱激活和缺乏结合表明2-ME可能通过间接机制调节PPAR-associated基因，可能涉及考克斯-2。事实上，在不抑制微管蛋白聚合的浓度下，2-ME的抗促有丝分裂作用被PPAR拮抗剂GW 9662和考克斯-2抑制剂NS 398阻断。最后，我们证明了2-ME抑制缺氧诱导因子-1 α。鉴定2-ME正或负调控的候选基因为研究和更好地理解2-ME诱导血管保护作用的机制提供了重要线索。
Methoxyestradiol (2-ME; estradiol metabolite) inhibits vascular smooth muscle cell (VSMC) growth and protects against atherosclerosis and vascular injury; however, the mechanisms by which 2-ME induces these actions remain obscure. To assess the impact of 2-ME on biochemical pathways regulating VSMC biology, we employed high-density oligonucleotide microarrays to identify differentially expressed genes in cultured human female aortic VSMCs treated with 2-ME acutely (4hrs) or long-term (30hrs). Both single gene analysis and Gene Set Enrichment Analysis (GSEA) revealed 2-ME-induced down-regulation of genes involved in mitotic spindle assembly and function in VSMCs. Also GSEA analysis identified effects of 2-ME on genes regulating cell-cycle progression, cell migration/adhesion, vasorelaxation, inflammation and cholesterol metabolism. Transcriptional changes were associated with changes in protein expression including inhibition of cyclin-D1, cyclin-B1, cdk6, cdk4, tubulin polymerization and cholesterol and steroid synthesis and up-regulation of cyclooxygenase-2 (COX-2) and matrix metalloproteinase-1. Microarray data suggested that 2-ME may activate peroxisome proliferator-activated receptors (PPARs) in VSMCs, and 2-ME has structural similarities with rosiglitazone (PPARγ agonist). However, our finding of weak activation and lack of binding of 2-ME to PPARs suggests that 2-ME may modulate PPAR-associated genes via indirect mechanisms, potentially involving COX-2. Indeed the anti-mitogenic effects of 2-ME at concentrations that do not inhibit tubulin polymerization were blocked by the PPAR antagonist GW9662 and COX-2 inhibitor NS398. Finally, we demonstrated that 2-ME inhibited hypoxia-inducible factor-1α. Identification of candidate genes that are positively or negatively regulated by 2-ME provides important leads to investigate and better understand the mechanisms by which 2-ME induces its vasoprotective actions.