Growth Of Bovine Aortic Endothelial Cells Research Articles

Anti-Angiogenic Effects of Conjugated Docosahexaenoic Acidin Vitroandin Vivo

The anti-angiogenic effects of conjugated docosahexaenoic acid (CDHA), which was prepared by an alkaline treatment of docosahexaenoic acid and contained conjugated double bonds, were investigated in vitro and in vivo. CDHA inhibited tube formation by the bovine aortic endothelial cell (BAEC), and also inhibited the proliferation of BAEC at a concentration of CDHA that suppressed tube formation, but did not influence cell migration. The inhibition of BAEC growth caused by CDHA was accompanied by a marked change in cellular morphology. Nuclear condensation and brightness were observed in Hoechst 33342-stained cells treated with CDHA, indicating that CDHA induced apoptosis in BAEC. We also evaluated the angiogenesis inhibition of CDHA in vivo. The vessel formation which was triggered by tumor cells was clearly suppressed in mice orally given CDHA. Our findings suggest that CDHA has potential use as a therapeutic dietary supplement for minimizing tumor angiogenesis.

Embellistatin, a microtubule polymerization inhibitor, inhibits angiogenesis both in vitro and in vivo

The efficient inhibition of angiogenesis is considered as a promising strategy for the treatment of angiogenesis-related diseases including cancer. Herein, we report that embellistatin, a bicyclic ketone compound known as a microtubule polymerization inhibitor, exhibits anti-angiogenic activity. Embellistatin inhibited in vitro angiogenesis of bovine aortic endothelial cells (BAECs) such as bFGF-induced invasion and tube formation as well as bFGF-induced mouse corneal angiogenesis in vivo. Notably, embellistatin exhibited stronger inhibition activity for the growth of BAECs than that of normal and cancer cell lines. Cell cycle analysis revealed that the compound arrests cell cycle at G 2/M phase, which is associated with the increased expression of p21 WAF1 and p53 partly. These results demonstrate that embellistatin may serve the basis for the development of new anti-angiogenic agents.

Synthesis of disaccharides derived from heparin and evaluation of effects on endothelial cell growth and on binding of heparin to FGF-2

The disaccharide β- d-GlcA-(1→4)-α- d-GlcNAc-1→OMe and other small nonsulfated oligosaccharides related to heparin/heparan sulfate have been shown to bind to FGF and activated the fibroblast growth factor (FGF) signalling pathway in (F32) cells expressing the FGF receptor. Synthetic routes to β- d-GlcA-(1→4)-α- d-GlcNAc-1→OMe and a glucose analogue β- d-Glc-(1→4)-α- d-GlcNAc-1→OMe are described. The effects of these disaccharides on endothelial cell growth, which is relevant to angiogenesis, were evaluated and it was found they did not mimic the inhibitory effects that were observed for heparin albumin (HA) and that have also been observed by monosaccharide conjugates. They did not alter bovine aortic endothelial cell (BAEC) proliferation, in the presence of FGF-2 in serum free medium or in absence of FGF-2 in serum free and complete medium. Disaccharides (10 μg/mL) reduced by 25–31% the inhibition caused by HA (10 μg/mL) on BAEC growth in serum-free medium but had no effect in complete medium. There was no evidence obtained for the binding of these oligosaccharides to FGF-2 in competition with HA by ELISA.

Synthesis of a glucuronic acid and glucose conjugate library and evaluation of effects on endothelial cell growth

Compounds that alter endothelial cell growth are of interest in the development of angiogenesis modulators. A structurally diverse series of saccharide derivatives (glycosylamide conjugates) have been synthesized and evaluated for their effects on bovine aortic endothelial cell (BAEC) growth. Heparin-albumin (HA) reduced BAEC growth by 32% at 10 μg/mL and a number of the novel saccharide conjugates from the library were found to mimic the effect of HA as they also inhibit endothelial cell survival under identical conditions. Two thiophene conjugates, thioglucamide (24% inhibition at 35 μM) and a related glucuronide (26% inhibition at 33 μM) were the most potent inhibitors of BAEC growth, as determined using a methylthiazol tetrazolium (MTT) assay. The effects of thioglucamide and HA on absolute cell number were also studied using cell counting experiments; thioglucamide (47% after 24 h) was more potent than indicated by the MTT assay and initially reduced the BAEC number to a greater extent than HA (30% after 24 h); however, its actions were over more rapidly than were HA's as cell growth had returned to levels of the control after 72 h where HA still caused 25% inhibition. The binding of the monosaccharide conjugates to fibroblast growth factor (FGF-2) in competition with heparin-albumin by ELISA was investigated to establish the possible mechanism by which glycoconjugates could alter growth but there was no general correlation between reduction in viable cell population and binding to FGF-2. No glycoconjugate reduced the proliferation of mouse mammary epithelial cells, nor did any alter gross cell morphology, supporting a proposal that the reduction in BAEC survival by monosaccharide conjugates such as thioglucamide is a result of the inhibition of cell proliferation rather than being an induction of cytotoxicity. These studies indicate that cell biological studies to determine the mechanism of action of the simple monosaccharide conjugates may be worthwhile.

Cochlioquinone A1, a New anti‐Angiogenic Agent from Bipolaris zeicola.

ChemInformVolume 35, Issue 9 Natural Products Cochlioquinone A1, a New anti-Angiogenic Agent from Bipolaris zeicola. Hye Jin Jung, Hye Jin Jung Dep. Biosci. Biotechnol., Sejong Univ., Seoul 143-747, S. KoreaSearch for more papers by this authorHyang Burm Lee, Hyang Burm Lee Dep. Biosci. Biotechnol., Sejong Univ., Seoul 143-747, S. KoreaSearch for more papers by this authorChi-Hwan Lim, Chi-Hwan Lim Dep. Biosci. Biotechnol., Sejong Univ., Seoul 143-747, S. KoreaSearch for more papers by this authorChang-Jin Kim, Chang-Jin Kim Dep. Biosci. Biotechnol., Sejong Univ., Seoul 143-747, S. KoreaSearch for more papers by this authorHo Jeong Kwon, Ho Jeong Kwon Dep. Biosci. Biotechnol., Sejong Univ., Seoul 143-747, S. KoreaSearch for more papers by this author Hye Jin Jung, Hye Jin Jung Dep. Biosci. Biotechnol., Sejong Univ., Seoul 143-747, S. KoreaSearch for more papers by this authorHyang Burm Lee, Hyang Burm Lee Dep. Biosci. Biotechnol., Sejong Univ., Seoul 143-747, S. KoreaSearch for more papers by this authorChi-Hwan Lim, Chi-Hwan Lim Dep. Biosci. Biotechnol., Sejong Univ., Seoul 143-747, S. KoreaSearch for more papers by this authorChang-Jin Kim, Chang-Jin Kim Dep. Biosci. Biotechnol., Sejong Univ., Seoul 143-747, S. KoreaSearch for more papers by this authorHo Jeong Kwon, Ho Jeong Kwon Dep. Biosci. Biotechnol., Sejong Univ., Seoul 143-747, S. KoreaSearch for more papers by this author First published: 03 February 2004 https://doi.org/10.1002/chin.200409230Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume35, Issue9March 2, 2004 RelatedInformation

Cochlioquinone A1, a new anti-angiogenic agent from Bipolaris zeicola

Cochlioquinone A1 (CoA1) was newly isolated from the culture extract of Bipolaris zeicola as a potent anti-angiogenic agent. CoA1 inhibited in vitro angiogenesis of bovine aortic endothelial cells (BAECs) such as bFGF-induced tube formation and invasion at the concentration (1 μg/mL) without cytotoxicity. Notably, CoA1 exhibited more potent inhibition activity for the growth of BAECs than that of normal and cancer cell lines investigated in this study. These results demonstrate that CoA1 is a new anti-angiogenic agent and can be developed as a new therapeutic agent for angiogenesis-related diseases.

High-level expression and secretion of recombinant mouse endostatin by Escherichia coli.

The expression of murine endostatin was achieved by placing its gene downstream of an alkaline phosphatase gene (phoA) promoter. To ensure proper folding and secretion of the recombinant protein, the mouse endostatin was fused with alkaline phosphatase signal peptide. SDS/polyacrylamide gel electrophoresis analysis of the culture medium of recombinant Escherichia coli cells revealed that endostatin was efficiently secreted. The signal peptide was efficiently cleaved during secretion as demonstrated by N-terminal amino acid sequencing. The maximum yield of secreted endostatin during fermentation was 40 mg/liter. Up to 28 mg of endostatin was purified from 1 liter of cell culture broth. The biological activity of recombinant protein was tested in a bovine aortic endothelial (BAE) cell proliferation assay. The recombinant endostatin inhibited the growth of BAE cells stimulated by basic fibroblast growth factor, and its ED50 was comparable to that from a previous report. Flow cytometric measurements of BAE cells cultivated in medium with endostatin demonstrated a cell cycle arrest mainly in the G0/G1 phase and a decrease in the S phase.

Overexpression of Protein Kinase C α and β1 Has Distinct Effects on Bovine Aortic Endothelial Cell Growth

Protein kinase C (PKC) plays an important role in the mitogenic response of endothelial cells to growth factors. PKC α and β1 are the predominant classical isoforms expressed by bovine aortic endothelial cells (BAECs). The present studies were undertaken to elucidate the effect of PKC α and β1 overexpression in BAEC growth. A series of BAEC lines that stably overexpress the full-length PKC α and β1 cDNA were generated by using a replication-defective recombinant retrovirus. The level of PKC α and β1 cDNA expression was determined by assaying for PKC α and β1 mRNA transcripts. PKC α and β1 protein levels were analysed by Western blotting. Functional analysis of these overexpressing lines was performed by measuring PKC activity and phorbol ester-binding assays. PKC α and β1 overexpression had distinctive effects on BAEC growth and cell-cycle progression. Relative to untransfected BAECs and BAECs transfected with the viral vector alone, BAECs that overproduced PKC α exhibited reduced proliferation in vitro and increased accumulation of cells in the G 2/M phase of the cell cycle. Growth inhibition was greater in cell lines overexpressing higher levels of PKC α. Conversely, a 5-fold greater increase in PKC β1 activity promoted BAEC growth and shortened BAEC doubling time, whereas cells with a 2- to 4-fold increase in enzyme activity had growth profiles similar to those of both control groups. These results suggest that PKC α and β1 overexpression has reciprocal effects on BAEC growth.

Vitamin E Restores the Glucose-Induced Impairment of Vascular Endothelial Cell Replication.

We evaluated the effect of vitamin E on the growth of cultured bovine aortic endothelial cells (BAEC). A high concentration of glucose (≥200mg/dl) in the culture medium reduced the replication of BAEC. Addition of vitamin E at a physiological concentration of 6μg/ml restored the impaired replication of BAEC cultured with a high concentration of glucose. Vitamin E had no effect on the growth of BAEC cultured with 100mg/dl glucose. Proliferation of BAEC was not significantly affected by exposure to mannitol at levels equimolar with a high concentration of glucose (600mg/dl), indicating that a high osmolality was not related to the glucose-induced reduction of replication of the cells. BAEC cultured in a high glucose medium may be susceptible to oxidative stress, and the benefit effect of vitamin E may be partly due to a reduction in oxidative damage. Vitamin E may protect endothelial cells against glucose-induced injury.

Radiofrequency plasma deposition of oxygen‐containing films on polystyrene and poly(ethylene terephthalate) substrates improves endothelial cell growth

Polystyrene and poly(ethylene terephthalate) substrates were modified by radiofrequency plasma deposition with organic vapors comprised of carbon, oxygen, and hydrogen (acetone, methanol, glutaraldehyde, formic acid, allyl alcohol, and ethylene oxide). The treatments resulted in the deposition of a film at least 100 A thick containing up to 26% atomic oxygen at the surface. A high oxygen incorporation was observed for vapors with a large oxygen-to-carbon ratio. Bovine aortic endothelial cell growth measured on acetone, methanol, and glutaraldehyde films was linearly correlated with the oxygen content of the treated surfaces. Nitrogen was incorporated in the surface by blending nitrogen gas into the organic vapor used for plasma deposition. The resulting nitrogen-containing substrates exhibited a high affinity for serum fibronectin but a moderate cell growth.

Ability of endothelial cells to condition culture medium.

Endothelial cell-conditioned medium contains two classes of factors distinguishable by behavior during dialysis and on specificity for cell type. One species, which diffuses through dialysis tubing with an exclusion limit of 6,000 to 8,000 daltons, supports growth of bovine aortic endothelial (BAE) cells in medium containing a growth limiting concentration of serum (0.2% serum). The production of this material appears to depend upon the presence of serum in the medium being conditioned. The activity increases with time of exposure of BAE cells to serum and with increasing concentration of serum present in the incubation medium. This activity cannot be replaced by exogenous epidermal growth factor, fibroblast growth factor, platelet-derived growth factor, insulin, or thymidine. The second species, the endothelial cell-derived growth factor (ECDGF), is retained by dialysis tubing with an exclusion limit of 6,000 to 8,000 daltons. ECDGF stimulates the growth of smooth muscle cells but does not support BAE cell growth in limiting serum concentrations. unlike the dialyzable species, the production of ECDGF is independent of previous incubation of BAE cell cultures in serum. These studies suggest that BAE cells are able to utilize serum components to produce conditioning factors for their own growth that are distinct from the higher molecular weight ECDGF.