Discovery of novel mechanosensitive genes in vivo using mouse carotid artery endothelium exposed to disturbed flow
Discovery of novel mechanosensitive genes in vivo using mouse carotid artery endothelium exposed to disturbed flow
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DOI：
10.1182/blood-2010-04-278192
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发表时间：
2010-10-14
期刊：
影响因子：
20.3
通讯作者：
Jo, Hanjoong
中科院分区：
文献类型：
--
作者：
Ni, Chih-Wen;Qiu, Haiwei;Jo, Hanjoong
最近，我们发现部分结扎小鼠颈动脉引起的血流紊乱会迅速诱发动脉粥样硬化。在这里，我们使用从血流扰动的左颈总动脉和未扰动的右颈总动脉分离的小鼠内皮RNA，通过全基因组微阵列研究鉴定了体内机械敏感基因。我们发现连接后 12 小时和 48 小时分别有 62 个和 523 个基因发生显着变化。结果通过定量聚合酶链式反应对 46 个测试基因中的 44 个进行了验证。这项阵列研究发现了许多新型机械敏感基因，包括 Lmo4、klk10 和 dhh，同时确认了众所周知的基因，如 Klf2、eNOS 和 BMP4。进一步验证了四个基因的蛋白质，包括 LMO4，它在小鼠主动脉弓和人冠状动脉内皮中以不对称模式表现出更高的表达。体内、离体和体外内皮基因表达谱的比较表明，许多体内机械敏感基因在培养过程中似乎丢失或失调。基因本体分析表明，扰动的流动在 12 小时内调节涉及细胞增殖和形态的基因，随后在 48 小时内调节炎症和免疫反应。确定这些新型机械敏感基因的功能重要性可能为了解血管生物学和动脉粥样硬化提供重要见解。 (血液.2010;116(15):e66-e73)
Recently, we showed that disturbed flow caused by a partial ligation of mouse carotid artery rapidly induces atherosclerosis. Here, we identified mechanosensitive genes in vivo through a genome-wide microarray study using mouse endothelial RNAs isolated from the flow-disturbed left and the undisturbed right common carotid artery. We found 62 and 523 genes that changed significantly by 12 hours and 48 hours after ligation, respectively. The results were validated by quantitative polymerase chain reaction for 44 of 46 tested genes. This array study discovered numerous novel mechanosensitive genes, including Lmo4, klk10, and dhh, while confirming well-known ones, such as Klf2, eNOS, and BMP4. Four genes were further validated for protein, including LMO4, which showed higher expression in mouse aortic arch and in human coronary endothelium in an asymmetric pattern. Comparison of in vivo, ex vivo, and in vitro endothelial gene expression profiles indicates that numerous in vivo mechanosensitive genes appear to be lost or dysregulated during culture. Gene ontology analyses show that disturbed flow regulates genes involved in cell proliferation and morphology by 12 hours, followed by inflammatory and immune responses by 48 hours. Determining the functional importance of these novel mechanosensitive genes may provide important insights into understanding vascular biology and atherosclerosis. (Blood.2010;116(15):e66-e73)