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Angiogenic actions of angiopoietin-1 require endothelium-derived nitric oxide

AMERICAN JOURNAL OF PATHOLOGY 2003年第6期162卷 1927-1936页

作者： Babaei, S Teichert-Kuliszewska, K Zhang, QW Jones, N Dumont, DJ Stewart, DJ Univ Toronto St Michaels Hosp Terrence Donnelly Heart Ctr Toronto ON M5B 1W8 Canada Univ Toronto St Michaels Hosp Div Cardiol Toronto ON M5B 1W8 Canada Univ Toronto Sunnybrook & Womens Coll Res Inst Div Mol & Cell Biol Res Toronto ON M5B 1W8 Canada Univ Toronto Dept Med Toronto ON M5B 1W8 Canada Univ Toronto Dept Med Biophys Toronto ON M5B 1W8 Canada

Angiopoietin1 (Ang1) is a novel angiogenic factor with important actions on endothelial cell (EC) differentiation and vascular maturation. Ang1 has been shown to prevent EC apoptosis through activation of PI3-kinase/ Akt, a pathway that is also known to activate endothelium nitric oxide synthase (eNOS). Therefore, we by pothesized that the angiogenic effects of Ang1 would also be dependent on the P13-kinase/Akt pathway, possibly mediated by increased eNOS activity and NO re lease. Treatment of human umbilical vein endothelial cells with recombinant Ang1* (300 ng/ml) for 15 minutes resulted in PI3-kinase-dependent Akt phosphorylation, comparable to that observed with vascular endothelial growth factor (VEGF) (50 ng/ml), and increased NO production in a PI3-kinase/Akt-dependent manner. Capillary-like tube formation induced by Ang1* in fibrin matrix at 24 hours (differentiation index, DI: 13.74 +/- 0.76 versus control 1.71 +/- 0.31) was abolished in the presence of the selective PI3-kinase inhibitor, LY294002 (50 mumol/L) (DI: 0.31 +/- 0.31, P < 0.01) or the NOS inhibitor, L-NAME (3 mmol/L) (DI: 4.10 +/- 0.59, P < 0.01). In subcutaneous Matrigel implants in vivo, addition of recombinant Ang1* or wild-type Ang1 from conditioned media of COS-1 cells transfected with a pFLAG Ang1 expression vector, induced significant neovascularization to a degree similar to VEGF. Finally, angiogenesis in vivo in response to both Ang1 and VEGF was significantly reduced in eNOS-deficient compared with wild-type mice. In summary, our results demonstrate for the first time that endothelial-derived NO is required for Ang1-induced angiogenesis, and that the PI3-kinase signaling mediates the activation of eNOS and NO release in response to Ang1. (Am J Pathol 2003,162:1927-1936)

关键词：雄二烯类/药理学血管生成诱导剂/遗传学血管生成诱导剂/代谢血管生成诱导剂/药理学血管生成素1 COS细胞细胞分化/药物作用细胞系色酮类/药理学剂量效应关系药物内皮血管/细胞学内皮血管/药物作用内皮血管/代谢酶抑制剂/药理学膜糖蛋白类/遗传学膜糖蛋白类/代谢膜糖蛋白类/药理学小鼠近交C57BL 小鼠基因敲除吗啉类/药理学 NG-硝基精氨酸甲酯/药理学新生血管化生理性/药物作用一氧化氮/代谢一氧化氮合酶/拮抗剂和抑制剂一氧化氮合酶/遗传学一氧化氮合酶/代谢磷酸肌醇3-激酶类/拮抗剂和抑制剂磷酰化/药物作用蛋白质丝氨酸苏氨酸激酶原癌基因蛋白质类/代谢原癌基因蛋白质c-akt 受体蛋白质酪氨酸激酶类/代谢受体 TIE-2 转染动物人类男(雄)性小鼠

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Internalization and processing of human angiogenin by cultured aortic smooth muscle cells

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 2000年第3期267卷 719-725页

作者： Hatzi, E Bassaglia, Y Badet, J Univ Paris 12 Inst Natl Sante & Rech Med URPESA 7053 Ctr Natl Rech SciLab Rech Croissance Cellulaire F-94010 Creteil France

Human angiogenin is a 14-kDa plasma protein with angiogenic and ribonucleolytic activities, Angiogenin binds specifically to aortic smooth muscle cells, activates second messenger pathways, and inhibits their proliferation, Human and bovine aortic smooth muscle cells were used to study the internalization and intracellular fate of human angiogenin at 37 degrees C, Using a specific antibody against angiogenin, we found that the internalized native protein was localized in the perinuclear region at 30 min and then dispersed throughout the cytoplasm. In conditions favoring receptor-mediated endocytosis, internalization of iodinated angiogenin showed a first peak at 5 min and then further increased for up to 24 h. The half-life of the molecule, calculated as 12 h in chase experiments, could contribute to its intracellular accumulation. In cell extracts, in addition to the 14-kDa protein, a 8.7-kDa fragment was observed at 24 h, and three fragments with molecular mass of 10.5, 8.7, and 6.1 kDa were detected at 48 h. Our data point to a specific internalization and processing of human angiogenin by aortic smooth muscle cells. (C) 1999 Academic Press.

关键词：血管生成诱导剂/代谢主动脉/细胞学主动脉/代谢生物转运细胞培养的细胞质/代谢半衰期动力学分子量肌平滑血管/细胞学肌平滑血管/代谢肽碎片/分析重组蛋白质类/代谢核糖核酸酶胰腺/代谢动物牛人类

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Cellular dysfunction in the diabetic fibroblast - Impairment in migration, vascular endothelial growth factor production, and response to hypoxia

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AMERICAN JOURNAL OF PATHOLOGY 2003年第1期162卷 303-312页

作者： Lerman, OZ Galiano, RD Armour, M Levine, JP Gurtner, GC NYU Med Ctr Inst Reconstruct Plast Surg Lab Microvasc Res & Vasc Tissue Engn New York NY 10016 USA

Although it is known that systemic diseases such as diabetes result in impaired wound healing, the mechanism for this impairment is not understood. Because fibroblasts are essential for wound repair, we compared the in vitro behavior of fibroblasts cultured from diabetic, leptin receptor-deficient (db/db) mice with wild-type fibroblasts from mice of the same genetic background in processes important during tissue repair. Adult diabetic mouse fibroblast migration exhibited a 75% reduction in migration compared to normal fibroblasts (P < 0.001) and was not significandy stimulated by hypoxia (1% O-2), whereas wildtype fibroblast migration was up-regulated nearly twofold in hypoxic conditions (P < 0.05). Diabetic fibroblasts produced twice the amount of pro-matrix metalloproteinase-9 as normal fibroblasts, as measured by both gelatin zymography and enzyme-linked immunosorbent assay (P < 0.05). Adult diabetic fibroblasts exhibited a sevenfold impairment In vascular endothelial growth factor (VEGF) production (4.5 +/- 1.3 pg/ml versus 34.8 +/- 3.3 pg/ml, P < 0.001) compared to wild-type fibroblasts. Moreover, wild-type fibroblast production of VEGF increased threefold in response to hypoxia, whereas diabetic fibroblast production of VEGF was not up-regulated in hypoxic conditions (P < 0.001). To address the question whether these differences resulted from chronic hyperglycemia or absence of the leptin receptor, fibroblasts were harvested from newborn db/db mice before the onset of diabetes (4 to 5 weeks old). These fibroblasts showed no impairments in VEGF production under basal or hypoxic conditions, confirming that the results from db/db fibroblasts in mature mice resulted from the diabetic state and were not because of alterations in the leptin-leptin receptor axis. Markers of cellular viability including proliferation and senescence were not significantly different between diabetic and wild-type fibroblasts. We conclude that, in vitro, diabetic fibroblasts show select

关键词：血管生成诱导剂/遗传学血管生成诱导剂/代谢动物新生细胞衰老/生理学细胞分裂/生理学细胞低氧/生理学细胞运动/生理学细胞培养的糖尿病实验性/遗传学糖尿病实验性/代谢糖尿病实验性/病理学疾病模型动物酶联免疫吸附测定成纤维细胞/代谢成纤维细胞/病理学明胶/代谢基质金属蛋白酶2/代谢基质金属蛋白酶9/代谢小鼠近交C57BL 小鼠突变型血管内皮生长因子A 动物女(雌)性小鼠

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Angiogenesis in head and neck squamous cell carcinoma

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CLINICAL OTOLARYNGOLOGY 2000年第3期25卷 169-180页

作者： Homer, JJ Greenman, J Stafford, ND Univ Hull Acad Surg Unit Dept Otolaryngol Head & Neck Surg Hull HU6 7RX N Humberside England

Tumour angiogenesis has recently attracted a great deal of attention as a critical part of oncogenesis and a necessary prerequisite for a malignant phenotype. Research into this process not only offers new insights into tumour biology but is also leading to the development of realistic novel and minimally toxic anti-tumour therapies. Various pro-angiogenic and anti-angiogenic cytokines and pathways have been characterized and their interrelationships are becoming increasingly complex as new findings are made. This article reviews the current understanding of tumour angiogenesis, the basic mechanisms involved and the more important and investigated pathways and proteins involved.

关键词：血管生成诱导剂/拮抗剂和抑制剂血管生成诱导剂/代谢毛细血管通透性/生理学癌鳞状细胞/代谢癌鳞状细胞/治疗内皮血管/代谢生长物质/代谢头颈部肿瘤/代谢头颈部肿瘤/治疗人类

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Novel expression of vascular endothelial growth factor receptor (VEGFR)-3 and VEGF-C on corneal dendritic cells

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AMERICAN JOURNAL OF PATHOLOGY 2003年第1期163卷 57-68页

作者： Hamrah, P Chen, L Zhang, Q Dana, MR Schepens Eye Res Inst Immunol Lab Boston MA USA Massachusetts Eye & Ear Infirm Boston MA 02114 USA Harvard Univ Sch Med Dept Ophthalmol Boston MA 02115 USA

Vascular endothelial growth factor-3 (VEGFR-3) plays a critical role in embryonic cardiovascular development and is thought to be expressed exclusively on the lymphatic endothelium, high endothelial venules, and rarely on adult vascular endothelium. Recent evidence also suggests expression of VEGFR-3 on some tumor-associated macrophages. We have studied the expression of VEGFR-3, its ligand VEGF-C and the co-receptor neuropilin-2, in normal and inflamed corneas and characterized the phenotype and distribution of VEGFR-3(+) cells. Our data demonstrate, for the first time, the expression of VEGFR-3 on corneal dendritic cells (DC) and its up-regulation in inflammation. VEGFR-3(+) DC are CD11c(+)CD45(+)CD11b(+), and are mostly major histocompatibility (MHC) class II(-)CD80(-)CD86(-), indicating immature DC of a monocytic lineage. During inflammation, there is rapid increase in the number of VEGFR-3(+) DC in the cornea associated with heightened membranous expression as compared to a mostly intracellular expression in uninflamed tissue. VEGFR-3(+) DC in normal corneas are VEGF-C- neuropilin-2(-), but express VEGF-C in inflammation. interestingly, similar cells are absent both in the normal and inflamed skin. These data demonstrate. for the first time, the expression of VEGFR-3 and VEGF-C on tissue DC, which implicate a novel potential relationship between lymphangiogenesis and leukocyte trafficking in the eye.

关键词：血管生成诱导剂/遗传学血管生成诱导剂/代谢细胞培养的角膜/细胞学角膜/代谢树突细胞/细胞学树突细胞/代谢炎症/代谢小鼠近交BALBC 小鼠近交C3H 小鼠近交C57BL 神经纤毛蛋白质2/代谢皮肤/细胞学皮肤/代谢血管内皮生长因子C 血管内皮生长因子受体3/遗传学血管内皮生长因子受体3/代谢动物人类男(雄)性小鼠

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Wielding more power over angiogenesis

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MOLECULAR MEDICINE TODAY 2000年第4期6卷 138-139页

作者： Habeck, M

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Angiopoietin expression in synovial membranes from patients with RA

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ANNALS OF THE RHEUMATIC DISEASES 2003年第9期62卷 911-911页

作者： Smith, MD Repatriat Gen Hosp Rheumatol Res Unit Daw Pk SA 5041 Australia

Sep 1, 2003 We thank Dr Smith for his interest in our study and for his comments about the differences between protein levels of angiopoietin-1 in RA synovium detected by immunohistochemistry and mRNA levels for angio... 详细信息

关键词：血管生成诱导剂/代谢血管生成素1 关节炎类风湿/代谢基因表达膜糖蛋白类/代谢 RNA 信使/遗传学滑膜/代谢人类

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