NO mediates mural cell recruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels
NO mediates mural cell recruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels
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DOI：
10.1172/jci24015
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发表时间：
2005-07-01
影响因子：
15.9
通讯作者：
Fukumura, D
中科院分区：
文献类型：
--
作者：
Kashiwagi, S;Izumi, Y;Fukumura, D
NO已被证明可介导血管生成，但其在血管形态发生和成熟中的作用尚不清楚。应用活体显微镜、组织学分析、α-平滑肌肌动蛋白和硫酸软骨素蛋白多糖4染色、微传感器NO测量和一氧化氮合酶(NOS)抑制剂，我们发现在B16小鼠黑色素瘤中，NO调节壁细胞覆盖以及血管分支和纵向伸展，但不调节血管的周向生长。NO敏感的荧光探针4，5-二氨基荧光素成像、一氧化氮合酶免疫组织化学染色，以及使用一氧化氮合酶基因缺陷小鼠的研究表明，血管内皮细胞中的eNOS是一氧化氮的主要来源，并诱导了这些效应。为了进一步剖析NO在壁细胞募集和血管形态发生中的作用，我们进行了一系列独立的分析。Transwell和琼脂糖下迁移分析表明，内皮细胞来源的NO诱导壁细胞前体向内皮细胞定向迁移。体内组织工程血管模型显示，NO在血管灌流前介导内皮细胞-壁细胞的相互作用，并诱导壁细胞重新聚集到血管新生血管，血管分支，以及血管的纵向延伸和随后的稳定。这些数据表明，内皮细胞来源的NO诱导壁细胞募集以及随后的血管新生血管的形态形成和稳定。
NO has been shown to mediate angiogenesis; however, its role in vessel morphogenesis and maturation is not known. Using intravital microscopy, histological analysis, a-smooth muscle actin and chondroitin sulfate proteoglycan 4 staining, microsensor NO measurements, and an NO synthase (NOS) inhibitor, we found that NO mediates mural cell coverage as well as vessel branching and longitudinal extension but not the circumferential growth of blood vessels in B16 murine melanomas. NO-sensitive fluorescent probe 4,5-diaminofluorescein imaging, NOS immunostaining, and the use of NOS-deficient mice revealed that eNOS in vascular endothelial cells is the predominant source of NO and induces these effects. To further dissect the role of NO in mural cell recruitment and vascular morphogenesis, we performed a series of independent analyses. Transwell and under-agarose migration assays demonstrated that endothelial cell-derived NO induces directional migration of mural cell precursors toward endothelial cells. An in vivo tissue-engineered blood vessel model revealed that NO mediates endothelial-mural cell interaction prior to vessel perfusion and also induces recruitment of mural cells to angiogenic vessels, vessel branching, and longitudinal extension and subsequent stabilization of the vessels. These data indicate that endothelial cell-derived NO induces mural cell recruitment as well as subsequent morphogenesis and stabilization of angiogenic vessels.