Angiopoietin-like 7 is an anti-angiogenic protein required to prevent vascularization of the cornea.
Angiopoietin-like 7 is an anti-angiogenic protein required to prevent vascularization of the cornea.
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DOI：
10.1371/journal.pone.0116838
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Amano S
中科院分区：
文献类型：
--
作者：
Toyono T;Usui T;Yokoo S;Taketani Y;Nakagawa S;Kuroda M;Yamagami S;Amano S
我们试图确定角膜角膜细胞中表达的抗血管生成分子，该分子负责维持角膜的无血管性。在无血清条件下，将人脐静脉内皮细胞（HUVEC）与人真皮成纤维细胞或人角膜基质细胞一起培养。通过用抗CD31（一种血管标记物）的抗体对培养物进行免疫染色来估计显示血管形成的区域。我们还进行了微阵列基因表达分析，并选择了一种分子，血管生成素样7（ANGPTL7）进行进一步的功能研究与角膜细胞和小鼠体内。在正常无血清培养基中显示血管形成的区域在HUVEC与角膜基质细胞共培养时比在相同条件下与真皮成纤维细胞共培养时明显更小。微阵列分析显示ANGPTL7在角膜细胞中的表达高于真皮成纤维细胞。在体外，通过使用特异性siRNA抑制ANGPTL7表达导致比用对照siRNA转染细胞更大的管形成，并且当向培养物中加入重组ANGPTL7蛋白时，管形成的这种增加被消除。在体内，将ANGPTL7 PshRNA基质内注射到小鼠的无血管角膜基质中导致血管生长。 ANGPTL7在角膜细胞中大量表达，在维持角膜无血管性和透明度中起主要作用。
We sought to identify the anti-angiogenic molecule expressed in corneal keratocytes that is responsible for maintaining the avascularity of the cornea. Human umbilical vein endothelial cells (HUVECs) were cultured with either human dermal fibroblasts or with human corneal keratocytes under serum-free conditions. The areas that exhibited blood vessel formation were estimated by immunostaining the cultures with an antitibody against CD31, a blood vessel marker. We also performed microarray gene-expression analysis and selected one molecule, angiopoietin-like 7 (ANGPTL7) for further functional studies conducted with the keratocytes and in vivo in mice. Areas showing blood vessel formation in normal serum-free medium were conditions were markedly smaller when HUVECs were co-cultured with corneal keratocytes than when they were co-cultured with the dermal fibroblasts under the same conditions. Microarray analysis revealed that ANGPTL7 expression was higher in keratocytes than in dermal fibroblasts. In vitro, inhibiting ANGPTL7 expression by using a specific siRNA led to greater tube formation than did the transfection of cells with a control siRNA, and this increase in tube formation was abolished when recombinant ANGPTL7 protein was added to the cultures. In vivo, intrastromal injections of an ANGPTL7 PshRNA into the avascular corneal stroma of mice resulted in the growth of blood vessels. ANGPTL7, which is abundantly expressed in keratocytes, plays a major role in maintaining corneal avascularity and transparency.
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