Department of Hand, Plastic and Reconstructive Surgery, University of Heidelberg, BG Trauma Center Ludwigshafen, Ludwig-Guttmann Str. 13, 67071, Ludwigshafen, Germany.
Institute of Human Genetics, Saarland University, Homburg-Saar, Germany.
J Transl Med. 2019 Jan 11;17(1):22. doi: 10.1186/s12967-019-1767-9.
Vascular shear stress promotes endothelial cell sprouting in vitro. The impact of hemodynamic forces on microRNA (miRNA) and gene expression within growing vascular networks in vivo, however, remain poorly investigated. Arteriovenous (AV) shunts are an established model for induction of neoangiogenesis in vivo and can serve as a tool for analysis of hemodynamic effects on miRNA and gene expression profiles over time.
AV shunts were microsurgically created in rats and explanted on postoperative days 5, 10 and 15. Neoangiogenesis was confirmed by histologic analysis and micro-computed tomography. MiRNA and gene expression profiles were determined in tissue specimens from AV shunts by microarray analysis and quantitative real-time polymerase chain reaction and compared with sham-operated veins by bioinformatics analysis. Changes in protein expression within AV shunt endothelial cells were determined by immunohistochemistry.
Samples from AV shunts exhibited a strong overexpression of proangiogenic cytokines, oxygenation-associated genes (HIF1A, HMOX1), and angiopoetic growth factors. Significant inverse correlations of the expressions of miR-223-3p, miR-130b-3p, miR-19b-3p, miR-449a-5p, and miR-511-3p which were up-regulated in AV shunts, and miR-27b-3p, miR-10b-5p, let-7b-5p, and let-7c-5p, which were down-regulated in AV shunts, with their predicted interacting targets C-X-C chemokine receptor 2 (CXCR2), interleukin-1 alpha (IL1A), ephrin receptor kinase 2 (EPHA2), synaptojanin-2 binding protein (SYNJ2BP), forkhead box C1 (FOXC1) were present. CXCL2 and IL1A overexpression in AV shunt endothelium was confirmed at the protein level by immunohistochemistry.
Our data indicate that flow-stimulated angiogenesis is determined by an upregulation of cytokines, oxygenation associated genes and miRNA-dependent regulation of FOXC1, EPHA2 and SYNJ2BP.
血管切变力促进体外内皮细胞发芽。然而，血流动力学对体内正在生长的血管网络中 microRNA（miRNA）和基因表达的影响仍未得到充分研究。动静脉（AV）分流术是体内诱导新生血管形成的一种成熟模型，可作为分析随时间推移血流动力学对 miRNA 和基因表达谱影响的工具。
通过显微外科手术在大鼠中创建 AV 分流术，并在术后第 5、10 和 15 天进行移植。通过组织学分析和微计算机断层扫描确认新生血管形成。通过微阵列分析和定量实时聚合酶链反应确定 AV 分流术组织标本中的 miRNA 和基因表达谱，并通过生物信息学分析与假手术静脉进行比较。通过免疫组织化学测定 AV 分流术内皮细胞内蛋白质表达的变化。
AV 分流术样本表现出强烈的促血管生成细胞因子、氧合相关基因（HIF1A、HMOX1）和血管生成生长因子的过度表达。miR-223-3p、miR-130b-3p、miR-19b-3p、miR-449a-5p 和 miR-511-3p 的表达显著负相关，这些 miRNA 在 AV 分流术中上调，而 miR-27b-3p、miR-10b-5p、let-7b-5p 和 let-7c-5p 在 AV 分流术中下调，其预测的相互作用靶点 C-X-C 趋化因子受体 2（CXCR2）、白细胞介素 1 阿尔法（IL1A）、表皮生长因子受体激酶 2（EPHA2）、突触结合蛋白 2 结合蛋白（SYNJ2BP）和叉头框 C1（FOXC1）存在。免疫组织化学证实 AV 分流术内皮细胞中 CXCL2 和 IL1A 的过度表达。
我们的数据表明，血流刺激的血管生成是由细胞因子的上调、氧合相关基因以及 miRNA 对 FOXC1、EPHA2 和 SYNJ2BP 的调节决定的。
