scholarly journals Taohong Siwu (TSW) decoction enhances angiogenesis by regulation of VHL/HIF-1α/VEGF pathway

2020 ◽  
Author(s):  
Zhi Tang ◽  
Wangyang Li ◽  
Hongzan Xie ◽  
Shengping Jiang ◽  
Yunqing Pu ◽  
...  
Keyword(s):  
Wound Healing ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Invasion ◽  
Bone Repair ◽  
Dependent Manner ◽  
Aortic Endothelial Cells ◽  
Vegf Pathway ◽  
Dose Dependent ◽  
Aortic Endothelial

Abstract Background: The incidence of bone fracture and bone-related diseases is increasing year by year. Angiogenesis plays vital role in fracture healing and bone repair. In this study, we assessed the effect of Taohong Siwu (TSW) decoction on angiogenesis by treatment of isolated rat aortic endothelial cell with TSW-containing serum. Method: First, TSW-containing serum was prepared from male Sprague-Dawley by intragastrically administration of TSW decoction. Then, isolated aortic endothelial cells were treated with different dose of TSW-containing serum. The effect of TSW-containing serum on the viability/proliferation of aortic endothelial cells were examined by MTT assay. The effects of TSW-containing serum on endothelial cell invasion, migration/spreading were detected by trans-well assay and scratch/wound-healing assay respectively. In addition, the effect of HIF-1α inhibitor on TSW-induced cell invasion and migration was also assessed. Moreover, the effect of TSW-containing serum on the expression of HIF-1α signaling pathway related protein, including HIF-1α, VHL and VEGF was detected by qRT-PCR and western-blot. Results: Our results showed that TSW decoction significantly increased the endothelial viability, invasion and wound-healing in a dose-dependent manner. Importantly, these enhanced effects induced by TSW were attenuated by HIF-1α inhibitor. Furthermore, our data demonstrated that TSW-containing serum increased the expression of HIF-1α and VEGF in a dose-dependent manner, while decreased the expression of VHL slightly. These effects induced by TSW were reversed by HIF-1α inhibitor. Conclusion: In summary these results for the first time suggested that TSW decoction enhances the angiogenesis by regulating of VHL/HIF-1α/VEGF pathway.

scholarly journals Taohong Siwu-Containing Serum Enhances Angiogenesis in Rat Aortic Endothelial Cells by Regulating the VHL/HIF-1α/VEGF Signaling Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Zhi Tang ◽  
Wangyang Li ◽  
Hongzan Xie ◽  
Shengping Jiang ◽  
Yunqing Pu ◽  
...  
Keyword(s):  
Wound Healing ◽  
Endothelial Cells ◽  
Signaling Pathway ◽  
Bone Repair ◽  
Dependent Manner ◽  
Sprague Dawley ◽  
Aortic Endothelial Cells ◽  
Vegf Signaling ◽  
Vegf Signaling Pathway ◽  
Aortic Endothelial

Background. The incidence of bone fracture and bone-related diseases is increasing every year. Angiogenesis plays a vital role in fracture healing and bone repair. This study assessed the benefits of Taohong Siwu (TSW) decoction on angiogenesis in isolated rat aortic endothelial cells (RAEC) treated with TSW-containing serum. Methods. The components of TSW decoction were analyzed by liquid chromatography-mass spectrometry (LC-MS). TSW-containing serum was prepared by gavage of TSW decoction to Sprague-Dawley (SD) rats. The effects of TSW-containing serum on the viability, migration, wound healing, and angiogenesis of RAEC were detected by the MTT, transwell, wound healing, and Matrigel lumen formation assays, respectively. In addition, the effects of an HIF-1α inhibitor on TSW-containing serum-induced RAEC were also assessed. The effects of TSW-containing serum on the expression of the HIF-1α signaling pathway were evaluated by qRT-PCR and western blot analysis. Results. LC-MS revealed that TSW decoction primarily contained isomaltulose, choline, D-gluconic acid, L-pipecolic acid, hypotaurine, albiflorin, and tryptophan. TSW-containing serum significantly increased the viability, migration, wound healing, and angiogenesis of RAEC in a dose-dependent manner. Furthermore, our results demonstrated that HIF-1α and VEGF expressions were increased in the cells of TSW-containing serum groups, whereas VHL expression was decreased. The effects of TSW-containing serum were reversed by treatment with an HIF-1α inhibitor. Conclusion. These results suggested that TSW decoction enhanced angiogenesis by regulating the VHL/HIF-1α/VEGF signaling pathway.

PMA-activated neutrophils decrease ectoenzyme activities in rabbit aortic endothelial cells in culture

1992 ◽  
Vol 263 (6) ◽  
pp. L657-L663
Author(s):  
X. Chen ◽  
M. Tzanela ◽  
M. K. Baumgartner ◽  
J. R. McCormick ◽  
J. D. Catravas
Keyword(s):  
Endothelial Cells ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Aortic Endothelial Cells ◽  
Endothelial Monolayers ◽  
Activated Neutrophils ◽  
Ace Activity ◽  
Dose Dependent Decrease ◽  
Dose Dependent ◽  
Aortic Endothelial

We have studied the effects of phorbol 12-myristate 13-acetate (PMA)-activated neutrophils [polymorphonuclear leukocytes (PMN)] on endothelial ectoenzyme [angiotensin-converting enzyme (ACE) and 5'-nucleotidase (NCT)] activities in cultured rabbit aortic endothelial cells (EC) with the use of [3H]benzoyl-Phe-Ala-Pro and 14C-labeled AMP as substrates, respectively, under first-order reaction conditions. PMA (1–1,000 ng/ml) or PMN alone had no effect on ACE activity. When PMA was incubated together with PMN (PMN/EC = 1.25:1 or 2.5 x 10(5) neutrophils/ml) for 4 h in Earle's salts, a PMA dose-dependent decrease in ACE activity was observed. Threshold PMA concentration was 2 ng/ml. At 8 ng PMA/ml, ACE activity was totally abolished, without any evidence of cytotoxicity, as inferred from release of 51Cr from prelabeled EC. The decrease in ACE activity was also dependent on PMN concentration and was detectable at PMN/EC values as low as 1.25:10 (0.25 x 10(5) PMN/ml). Inhibition of ACE occurred as early as 1 h after incubation (PMA 10 ng/ml, PMN/EC = 1.25:1). PMA alone caused a small but significant increase in NCT activity, whereas PMA coincubation with PMN produced a significant decrease in NCT activity (20–29%), which was PMA and PMN concentration independent. PMA increased PMN adherence to endothelial monolayers in a concentration-dependent manner. Pretreating PMN with monoclonal antibody 60.3 (raised against the adhesion glycoprotein CD18) or placing a 2-microns filter between PMN and EC, protected the decrease in ACE activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of DL-Penicillamine on Cytotoxic Reaction between Baboon Performed Xenoantibodies and Pig Endothelial Cells

1997 ◽  
Vol 20 (7) ◽  
pp. 375-378
Author(s):  
P.A. Kwiatkowski ◽  
J. Puc ◽  
W. Rowinski ◽  
P. Fiedor
Keyword(s):  
Endothelial Cells ◽  
Cytotoxicity Assay ◽  
Dependent Manner ◽  
Elisa Method ◽  
Aortic Endothelial Cells ◽  
Cytotoxic Reaction ◽  
Igg Binding ◽  
Dose Dependent ◽  
Aortic Endothelial

The purpose of this study was to evaluate effects of DL-Penicillamine (DLP), a compound interrupting S-S bonds (IgM pentamers) on binding and cytotoxicity of adult baboon performed xenoantibodies to pig endothelial cells. Pooled baboon serum was treated with different concentrations of DLP during various periods of time. Complement-mediated cytotoxicity assay was used to determine the reactivity of baboon xenoantibodies to pig aortic endothelial cells (PAEC). To assess IgM and IgG binding to PAEC, ELISA method was applied. Serum treated with DLP revealed significant reduction of cytotoxicity in a dose dependent manner. Cytotoxicity was also reduced during time prolongation of DLP exposure to PAEC. Results indicate that baboon performed IgM and IgG xenoantibodies bind to pig endothelial cells, but only IgM is able to cause degradation of the complement. DLP significantly reduces cytotoxicity and eliminates binding of IgMs to PAEC in spite of continued binding of IgG xenoantibodies to the surface of endothelium.

Purpurogallin protects both ventricular myocytes and aortic endothelial cells of rats against oxyradical damage

1992 ◽  
Vol 70 (9) ◽  
pp. 803-809 ◽  
Author(s):  
Tai-Wing Wu ◽  
Jun Wu ◽  
Doug Carey ◽  
Ling-Hua Zeng
Keyword(s):  
Endothelial Cells ◽  
Xanthine Oxidase ◽  
Ventricular Myocytes ◽  
Cell Damage ◽  
Dependent Manner ◽  
Aortic Endothelial Cells ◽  
Electron Microscopies ◽  
First Time ◽  
Dose Dependent ◽  
Aortic Endothelial

Rat ventricular myocytes have been isolated and cultured by two separate procedures. Using phase-contrast and electron microscopies, we illustrate that (a) definitive cell damage is produced when myocytes are exposed to xanthine oxidase – hypoxanthine and (b) purpurogallin between 0.25 and 1.0 mM prolongs survival of both myocyte preparations in a dose-dependent manner. The cytoprotection produced by 1 mM purpurogallin exceeds that given by 2 mM each of ascorbate, Trolox, and mannitol, or 24 200 IU superoxide dismutase/L and (or) 92 000 IU catalase/L. Furthermore, we noted, for the first time, that purpurogallin markedly protects rat aortic endothelial cells, a key target of free radical generation and attack. In contrast, Trolox has a negligible effect here. Mechanistically, we showed that purpurogallin inhibits urate formation by xanthine oxidase more potently than allopurinol. Also, the compound diminishes formation of superoxide-reduced cytochrome c. Therefore, purpurogallin is a potent protector of ventricular myocytes and aortic endothelial cells, both of which are important cells in the cardiovascular system.Key words: purpurogallin, endothelial cells, myocytes.

Molecular characterization of the Na-K-Cl cotransporter of bovine aortic endothelial cells

1997 ◽  
Vol 273 (1) ◽  
pp. C188-C197 ◽  
Author(s):  
T. R. Yerby ◽  
C. R. Vibat ◽  
D. Sun ◽  
J. A. Payne ◽  
M. E. O'Donnell
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Amino Acid ◽  
Blot Analysis ◽  
Cdna Probe ◽  
Bovine Aortic Endothelial Cell ◽  
Coding Region ◽  
Cerebral Microvessels ◽  
Aortic Endothelial Cells ◽  
Aortic Endothelial

The Na-K-Cl cotransporter is an important regulator of endothelial cell volume and may also contribute to flux of Na and Cl across the endothelium of the blood-brain barrier. To date, two Na-K-Cl cotransport isoforms have been identified, the cotransporter in secretory epithelia, NKCC1, and that in absorptive renal epithelia, NKCC2. Our previous studies showed that a monoclonal antibody to the cotransporter of human colonic T84 epithelial cells, an NKCC1 isoform, recognizes a 170-kDa glycoprotein from endothelial cells. The molecular identity of the Na-K-Cl cotransporter present in endothelial cells, however, has been unknown. In addition, although evidence has been provided that phosphorylation of the endothelial cotransporter plays a role in regulating its activity, little is known about potential sites for protein kinase interaction with the cotransporter. The present study was conducted to determine the molecular structure of the endothelial Na-K-Cl cotransporter. Using a 1.0-kilobase (kb) cDNA fragment from a conserved region of the T84 cell cotransporter, we screened a bovine aortic endothelial cell cDNA library and subsequently identified and sequenced two overlapping clones that together spanned the entire coding region. The endothelial cotransporter is a 1,201-amino acid protein with 12 putative transmembrane segments and large amino and carboxy termini, each containing several consensus sites for phosphorylation by protein kinases. Comparison of the endothelial cotransporter amino acid sequence with known NKCC1 and NKCC2 sequences revealed a 96% identity with NKCC1. Northern blot analysis using a cDNA probe from the endothelial cotransporter revealed high expression of approximately 7.5-kb transcripts in a number of bovine tissues. Finally, a prominent expression of Na-K-Cl cotransporter was found by Western blot analysis in both cultured and freshly isolated endothelial cells of bovine aorta and cerebral microvessels.

scholarly journals Clinopodium tomentosum (Kunth) Govaerts Leaf Extract Influences in vitro Cell Proliferation and Angiogenesis on Primary Cultures of Porcine Aortic Endothelial Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-11
Author(s):  
Irvin Tubon ◽  
Chiara Bernardini ◽  
Fabiana Antognoni ◽  
Roberto Mandrioli ◽  
Giulia Potente ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Primary Cultures ◽  
Ethanolic Extract ◽  
Total Phenolic ◽  
Dependent Manner ◽  
Aortic Endothelial Cells ◽  
Phenolic Components ◽  
Porcine Aortic Endothelial Cells ◽  
Aortic Endothelial

Clinopodium tomentosum (Kunth) Govaerts is an endemic species in Ecuador, where it is used as an anti-inflammatory plant to treat respiratory and digestive affections. In this work, effects of a Clinopodium tomentosum ethanolic extract (CTEE), prepared from aerial parts of the plant, were investigated on vascular endothelium functions. In particularly, angiogenesis activity was evaluated, using primary cultures of porcine aortic endothelial cells (pAECs). Cells were cultured for 24 h in the presence of CTEE different concentrations (10, 25, 50, and 100 μg/ml); no viability alterations were found in the 10-50 μg/ml range, while a slight, but significant, proliferative effect was observed at the highest dose. In addition, treatment with CTEE was able to rescue LPS-induced injury in terms of cell viability. The CTEE ability to affect angiogenesis was evaluated by scratch test analysis and by an in vitro capillary-like network assay. Treatment with 25-50 μg/ml of extract caused a significant increase in pAEC’s migration and tube formation capabilities compared to untreated cells, as results from the increased master junctions’ number. On the other hand, CTEE at 100 μg/ml did not induce the same effects. Quantitative PCR data demonstrated that FLK-1 mRNA expression significantly increased at a CTEE dose of 25 μg/ml. The CTEE phytochemical composition was assessed through HPLC-DAD; rosmarinic acid among phenolic acids and hesperidin among flavonoids were found as major phenolic components. Total phenolic content and total flavonoid content assays showed that flavonoids are the most abundant class of polyphenols. The CTEE antioxidant activity was also showed by means of the DPPH and ORAC assays. Results indicate that CTEE possesses an angiogenic capacity in a dose-dependent manner; this represents an initial step in elucidating the mechanism of the therapeutic use of the plant.

scholarly journals Haemophilus somnus Induces Apoptosis in Bovine Endothelial Cells In Vitro

2001 ◽  
Vol 69 (3) ◽  
pp. 1650-1660 ◽  
Author(s):  
Matt J. Sylte ◽  
Lynette B. Corbeil ◽  
Thomas J. Inzana ◽  
Charles J. Czuprynski
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Apoptosis ◽  
Dependent Manner ◽  
Endothelial Cell Apoptosis ◽  
Heat Stable ◽  
Culture Filtrates ◽  
Haemophilus Somnus ◽  
Bovine Endothelial Cells ◽  
Dose Dependent

ABSTRACT Haemophilus somnus causes pneumonia, reproductive failure, infectious myocarditis, thrombotic meningoencephalitis, and other diseases in cattle. Although vasculitis is commonly seen as a result of systemic H. somnus infections, the pathogenesis of vascular damage is poorly characterized. In this study, we demonstrated that H. somnus (pathogenic isolates 649, 2336, and 8025 and asymptomatic carrier isolates 127P and 129Pt) induce apoptosis of bovine endothelial cells in a time- and dose-dependent manner, as determined by Hoechst 33342 staining, terminal deoxynucleotidyl transferase-mediated dUTP-FITC nick end labeling, DNA fragmentation, and transmission electron microscopy. H. somnus induced endothelial cell apoptosis in as little as 1 h of incubation and did not require extracellular growth of the bacteria. Viable H. somnus organisms induced greater endothelial cell apoptosis than heat-killed organisms. Since viableH. somnus cells release membrane fibrils and blebs, which contain lipooligosaccharide (LOS) and immunoglobulin binding proteins, we examined culture filtrates for their ability to induce endothelial cell apoptosis. Culture filtrates induced similar levels of endothelial cell apoptosis, as did viable H. somnus organisms. Heat inactivation of H. somnus culture filtrates partially reduced the apoptotic effect on endothelial cells, which suggested the presence of both heat-labile and heat-stable factors. We found thatH. somnus LOS, which is heat stable, induced endothelial cell apoptosis in a time- and dose-dependent manner and was inhibited by the addition of polymyxin B. These data demonstrate that H. somnus and its LOS induce endothelial cell apoptosis, which may play a role in producing vasculitis in vivo.

scholarly journals Endothelial Cell Culture Model (ECCM) for Evaluation of Aortic Endothelial Cells Exposed to Normal and Disturbed Flow

2011 ◽  
Vol 17 (S2) ◽  
pp. 1218-1219
Author(s):  
R Estrada ◽  
G Giridharan ◽  
S Prabhu ◽  
P Sethu
Keyword(s):  
Cell Culture ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Culture Model ◽  
Endothelial Cell Culture ◽  
Disturbed Flow ◽  
Aortic Endothelial Cells ◽  
Culture Model ◽  
Aortic Endothelial

Extended abstract of a paper presented at Microscopy and Microanalysis 2011 in Nashville, Tennessee, USA, August 7–August 11, 2011.

Angiostatin and plasminogen share binding to endothelial cell surface actin

2005 ◽  
Vol 83 (1) ◽  
pp. 28-35 ◽  
Author(s):  
A K Dudani ◽  
M Ben-Tchavtchavadze ◽  
S Porter ◽  
E Tackaberry
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Surface ◽  
Dependent Manner ◽  
Proteolytic Fragment ◽  
Binding Motifs ◽  
Fluorescence Studies ◽  
Endothelial Cell Surface ◽  
Dose Dependent ◽  
Dependent Processes

Previous studies from this laboratory have demonstrated that plasminogen binds to endothelial cell surface-associated actin via its kringles in a dose-dependent and specific manner. The purpose of this study was to determine whether angiostatin, a proteolytic fragment of plasminogen, shares binding properties with plasminogen. Our results indicated that like plasminogen, angiostatin bound to actin in a time-, concentration-, and kringle-dependent manner. Furthermore, this binding was significantly inhibited by excess plasminogen, suggesting that both proteins shared binding motifs on the actin molecule. Fluorescence studies demonstrated that angiostatin bound to intact endothelial cells through its kringles, and this binding was also inhibited by plasminogen but not by unrelated proteins. Ligand blot analyses on endothelial cell lysates indicated that angiostatin interacted with a 42 kDa protein, which was identified as actin. Furthermore, an anti-actin antibody inhibited binding of angiostatin to endothelial cells by approximately 25%. These results suggest that angiostatin and plasminogen share binding to endothelial cell surface actin and, therefore, that angiostatin has the potential to inhibit plasmin-dependent processes such as cell migration–movement.Key words: plasminogen, angiostatin, endothelial cells, actin.

scholarly journals Tumor necrosis factor induction of endothelial cell urokinase-type plasminogen activator mediated proteolysis of extracellular matrix and its antagonism by gamma-interferon

Blood ◽  
1992 ◽  
Vol 79 (3) ◽  
pp. 678-687 ◽  
Author(s):  
MJ Niedbala ◽  
MS Picarella
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Plasminogen Activator ◽  
Inhibitory Response ◽  
Dependent Manner ◽  
Ecm Degradation ◽  
Ifn Treatment ◽  
Dose Dependent ◽  
Necrosis Factor ◽  
Pai 1

Tumor necrosis factor (TNF) has a profound capacity to alter the endothelial cell phenotype that includes morphologic and functional changes that may be central for proinflammatory processes. Recent observations have indicated that TNF can promote the synthesis and secretion of urokinase plasminogen activator (uPA) in low passage human endothelial cells that normally release little uPA. In this report we have confirmed and expanded upon these initial observations in human endothelial cells and describe the ability of gamma-interferon (gamma- IFN) to inhibit TNF-induced uPA synthesis and secretion in a dose- dependent manner (0.025 to 25 ng/mL). Analysis of cell-free conditioned medium derived from gamma-IFN-treated cultures by micro-enzyme-linked immunosorbent assay (ELISA) methodologies using uPA- and plasminogen activator inhibitor type 1 (PAI-1)-specific monoclonal antibodies (MoAbs) indicate that the decrease in uPA activity observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) zymography is a direct result of a decrease in extracellular uPA antigen and is not a consequence of increased PAI-1 antigen. These findings are supported by Northern blot analyses that indicate that gamma-IFN treatment of endothelial cells resulted in a decreased steady state level of uPA messenger RNA (mRNA) with no measurable change in PAI-1 mRNA. This inhibitory response is specific for gamma-IFN because alpha-IFN fails to elicit a similar inhibitory response. In addition, TNF augmented extracellular proteolysis of radiolabeled subendothelial extracellular matrix (ECM) in a dose-dependent manner. The observed increase in ECM degradation mediated by TNF treatment of endothelial cells was dependent on the presence of plasminogen and could be inhibited by an anticatalytic uPA MoAb implying the requirement of proteolytically active uPA in this process. gamma-IFN (25 ng/mL) treatment of endothelial cells antagonized TNF-promoted degradation of radiolabeled ECM at a concentration that completely inhibited TNF- mediated uPA expression and activity. In addition, endothelial cells treated with TNF (18 hours) displayed the ability to invade ECM and reorganize individual cells into tube-like structures that were not evident in untreated control cultures when grown on Matrigel-coated culture dishes. gamma-IFN treatment of endothelial cells propagated on Matrigel was observed to inhibit TNF-mediated ECM invasion and tube formation at concentrations that were analogous to those required for the inhibition of uPA expression and activity. In summary, these observations suggest a novel homeostatic control mechanism for endothelial cell regulation of subendothelial ECM degradation promoted by TNF and inhibited by gamma-IFN.(ABSTRACT TRUNCATED AT 400 WORDS).

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