Cerebral microvascular endothelial cell tube formation: role of astrocytic epoxyeicosatrienoic acid release

Munzenmaier, Diane H.; Harder, David R.

doi:10.1152/ajpheart.2000.278.4.h1163

Cited by 124 publications

(113 citation statements)

References 28 publications

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“…Cerebral angiogenesis is an intricate process that differs greatly from the analogous process in non-neural tissues. A significant difference is the presence and involvement of astrocytes in angiogenesis within the nerve system (22,23). Although it is currently unknown whether astrocytes derived from NSCs stimulate vascular tube formation in this model, this in vitro 3-D culture system of NSCs appears to mimic cerebral vascular development.…”

Section: Discussionmentioning

confidence: 99%

Angiogenesis In Vitro: Vascular Tube Formation From the Differentiation of Neural Stem Cells

et al. 2004

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Abstract. Neural stem cells (NSCs) were isolated from the mouse cortex on embryonic day 12.5 and cultured by neurosphere formation in serum-free medium in the presence of basic fibroblast growth factor (bFGF). When NSCs were inoculated in collagen gels with 10% fetal bovine serum (FBS) and bFGF and incubated for 10 days, vessel-like tube structures consisting of PECAM-1-or VE-cadherin-immunoreactive cells were formed in the gels. Moreover, the formation of vascular tube-like structures with a massive investment of a -smooth muscle actin-immunoreactive or GFAP-immunoreactive cells was occasionally observed, indicating angiogenesis identical to cerebral vascular development in vivo. To examine whether NSCs are capable of producing endothelial cells, differentiation was induced by the addition of 10% FBS after bFGF withdrawal. Most of the cells displayed a cobblestone-like morphology. Immunological analyses and RT-PCR indicated that NSCs expressed endothelial cell-specific marker proteins such as PECAM-1, VE-cadherin, and Flk-1; and these expressions were maintained or up-regulated during differentiation. Similar tube structures were also observed when the differentiated cells were inoculated in collagen gels and incubated for 5 days. These results suggested that NSCs give rise to two types of vascular cells, endothelial cells and mural cells in vitro, which have the ability to form vascular tubes.

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Section: Discussionmentioning

confidence: 99%

Angiogenesis In Vitro: Vascular Tube Formation From the Differentiation of Neural Stem Cells

et al. 2004

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“…In endothelial cells, CYP 2C9 overexpression and 11,12-EET application induce proliferation and angiogenesis (Michaelis et al, 2003) and an angiogenic effect of EETs derived from astrocytes has also been described (Munzenmaier and Harder, 2000;Zhang and Harder, 2002). As the process of angiogenesis involves endothelial cell migration and the degradation of the extracellular matrix in addition to cell proliferation, we assessed the effects of CYP 2C upregulation on these processes.…”

Section: Discussionmentioning

confidence: 99%

Cytochrome P450 epoxygenases 2C8 and 2C9 are implicated in hypoxia-induced endothelial cell migration and angiogenesis

Michaelis

Fißlthaler

Barbosa-Sicard

et al. 2005

Journal of Cell Science

146

122

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Recent studies suggest that cytochrome P450 (CYP) epoxygenase-derived epoxyeicosatrienoic acids (EETs) elicit cell proliferation and promote angiogenesis. The aim of this study was to determine the role of CYP 2C8/9-derived EETs in the process of angiogenesis under hypoxic conditions. In human endothelial cells, hypoxia enhanced the activity of the CYP 2C9 promoter, increased the expression of CYP 2C mRNA and protein and augmented 11,12-EET production. In Transwell assays, the migration of endothelial cells pre-exposed to hypoxia to increase CYP expression was abolished by CYP 2C antisense oligonucleotides as well as by the CYP inhibitor MS-PPOH and the EET antagonist 14,15-epoxyeicosa-5(Z)-enoic acid (EEZE). Similar findings were obtained in porcine coronary artery endothelial cells. CYP 2C9 overexpression in endothelial cells increased the association of PAK-1 with Rac, a response also elicited by the CYP 2C9 product 11,12-EET. Matrix metalloprotease (MMP) activity was increased in CYP-2C9-overexpressing cells and correlated with increased invasion through Matrigel-coated Transwell chambers: an effect sensitive to the CYP 2C9 inhibitor sulfaphenazole as well as to EEZE and the MMP inhibitor GM6001. In in vitro angiogenesis models, the EET antagonist inhibited tube formation induced by CYP 2C9 overexpression as well as that in endothelial cells exposed to hypoxia to increase CYP 2C expression. Furthermore, in the chick chorioallantoic membrane assay, EEZE abolished hypoxia-induced angiogenesis. Taken together, these data indicate that CYP 2C-derived EETs significantly affect the sequence of angiogenic events under hypoxic conditions. Journal of Cell Science 5490 physiological/pathophysiological response. The aim of the present investigation was to determine the effects of a potent angiogenic stimulus (hypoxia) on the expression of CYP 2C protein in endothelial cells and to further elucidate the mechanisms by which CYP 2C-derived EETs promote the degradation of the extracellular matrix and endothelial cell migration: two essential steps in the process of angiogenesis. Materials and Methods MaterialsMatrigel was from BD Biosciences, 11,12-EET was purchased from Cayman Chemicals (Massy, France), KT 5720 and AG 1478 were from Calbiochem (Darmstadt, Germany) and thrombin was from Haemochrom Diagnostica GmbH (Essen, Germany). MS-PPOH and 14,15-epoxyeicosa-5(Z)-enoic acid (14,15-EEZE) were synthesized as described (Gauthier et al., 2002). Sulfaphenazole, myelin basic protein, the antibody recognizing ␤-actin and all other chemicals were from Sigma. Cell cultureHuman umbilical vein endothelial cells (HUVEC) were either isolated as described (Busse and Lamontagne, 1991) or purchased from Cell Systems/Clonetics (Solingen, Germany). Cells were cultured in MCDB 131 (Gibco Life Technology, Karlsruhe, Germany), supplemented with 8% fetal calf serum (FCS), L-glutamine (10 mmol/l), basic fibroblast growth factor (1 ng/ml), epidermal growth factor (0.1 ng/ml), endothelial cell growth-stimulating factor from bovine brain (ECGS/H...

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“…These nonvasodilatory activities affect the function and viability of vascular endothelial cells and include up-regulation of eNOS at the levels of mRNA, protein, posttranslational modification, protection from apoptosis induced by tumor necrosis factor-␣, and reduction of expression of cytokine-induced endothelial cell adhesion molecules, thereby preventing leukocyte adhesion to the vascular wall (Fisslthaler et al, 1999;Node et al, 1999;Wang et al, 2003a,b). Recently, astrocyte-derived EETs were shown to induce cerebral capillary endothelial cell mitogenesis and tube formation (Munzenmaier and Harder, 2000), and adenovirus-mediated CYP2C9 gene transfection and exogenous 14,15-EET were shown to exert similar effects in human lung endothelial cell lines (Medhora et al, 2003). In addition, CYP2C9 overexpression in endothelial cells was shown to induce endothelial tube formation via stimulating cyclooxygenase-2 expression and prostacyclin production (Michaelis et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

Arachidonic Acid Epoxygenase Metabolites Stimulate Endothelial Cell Growth and Angiogenesis via Mitogen-Activated Protein Kinase and Phosphatidylinositol 3-Kinase/Akt Signaling Pathways

Wang

Wei

Xiao

et al. 2005

J Pharmacol Exp Ther

178

134

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Cytochrome P450 arachidonic acid (AA) epoxygenase metabolites, the epoxyeicosatrienoic acids (EETs), dilate arteries via hyperpolarization of smooth muscle cells and also have nonvasodilatory effects within the vasculature. The present study investigated the angiogenic effects of endogenous and exogenous EETs and the relevant signaling mechanisms involved. Bovine aortic endothelial cells (BAECs) were incubated with synthetic EETs or infected with recombinant adeno-associated viruses (rAAVs) containing CYP2C11-NADPH-cytochrome P450 oxidoreductase (CYPOR), CYP2J2, or CYP102 F87V mutant to increase endogenous levels of EETs. The following endpoints were measured: BAEC proliferation, migration, capillary formation, and in vivo angiogenesis. The potential involvement of various signaling pathways was explored using selective inhibitors. The results showed that transfection with either rAAV-CYP2C11-CYPOR, rAAV-CYP2J2, or rAAV-CYP102 F87V, or incubation with EETs promoted BAEC proliferation, increased migration of BAECs as assessed by Transwell analysis and wound healing assays, and enhanced capillary tubule formation as determined by chicken embryo chorioallantoic membrane assays and tube formation tests on matrigel. The effects of EETs on proliferation, migration, and capillary tubule formation were attenuated by inhibitors of mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3 (PI3)-kinase/ Akt pathways and partially attenuated by an endothelial nitricoxide synthase (eNOS) inhibitor but not by a protein kinase C inhibitor. In a rat ischemic hind limb model, rAAV-mediated AA epoxygenase transfection induced angiogenesis. We conclude that AA epoxygenase metabolites can promote angiogenesis, which may provide protection to ischemic tissues. The results also suggest that the angiogenic effects of EETs involve the MAPK and PI3-kinase/Akt signaling pathways, and to some extent, the eNOS pathway.Arachidonic acid (AA) is esterified to cell membrane glycerophospholipids and liberated by phospholipase A 2 in response to various stimuli. AA can be activated by three different enzyme pathways: the cyclooxygenase, the lipoxygenase, and the cytochrome P450 (P450) monooxygenase

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Cerebral microvascular endothelial cell tube formation: role of astrocytic epoxyeicosatrienoic acid release

Cited by 124 publications

References 28 publications

Angiogenesis In Vitro: Vascular Tube Formation From the Differentiation of Neural Stem Cells

Angiogenesis In Vitro: Vascular Tube Formation From the Differentiation of Neural Stem Cells

Cytochrome P450 epoxygenases 2C8 and 2C9 are implicated in hypoxia-induced endothelial cell migration and angiogenesis

Arachidonic Acid Epoxygenase Metabolites Stimulate Endothelial Cell Growth and Angiogenesis via Mitogen-Activated Protein Kinase and Phosphatidylinositol 3-Kinase/Akt Signaling Pathways

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