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

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)

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Stimulation Of Prostacyclin Production In Aortic Endothelial Cells By A Non-Dialysable Serum Factor

10.1055/s-0038-1651998 ◽

1981 ◽

Author(s):

E R Hall ◽

M Rafelson ◽

K Wu

Keyword(s):

Endothelial Cells ◽

Arachidonic Acid ◽

Calf Serum ◽

Dependent Manner ◽

Freezing And Thawing ◽

Concentration Dependent Manner ◽

Aortic Endothelial Cells ◽

Prostacyclin Production ◽

Aortic Endothelial

The production of prostacyclin (PGI2) by vascular endothelial cells is thought to be of primary importance in maintaining normal hemostasis. We have investigated the production of prostacyclin in bovine arterial endothelial cells maintained in Dulbecco’s modified Eagle’s medium (DMEM) containing 30% fetal calf serum. Intact, confluent monolayers of endothelial cells (3x106 cells) in passages 2 through 6 were used. The growth medium was removed and the cells were washed in DMEM that did not contain serum. 3 mls of medium alone or containing normal plasma or serum was then added and incubated at 37°C for 15 min. Then, 1 mg of arachidonic acid was added and the cells incubated for an additional hour. The test medium was removed, centrifuged to remove any loose cells and stored at -70°C. To determine the production of PGI2 by the endothelial cells, the medium was assayed for 6-keto-PGF1α, the stable metabolite of PGI2, by radioimmunoassay. The synthesis of prostacyclin by bovine aortic endothelial cells was significantly increased in a concentration dependent manner by both normal platelet poor plasma and normal serum. This increase in prostacyclin production was inhibited by both aspirin and indomethacin, indicating an increase in synthesis rather than the release of PGI2. Furthermore, this increase could be demonstrated in the presence or absence of added arachidonic acid. The active component in plasma and serum was non-dialysable, eliminating the possibility of a small compound such as bradykinin or angiotensin II. This active factor was present after freezing and thawing the plasma and serum and was heat stable (60°C, 5 min). The presence of an endogenous prostacyclin stimulating factor may be significant in the in vivo regulation of prostacyclin production.

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Permeability of endothelial monolayers to albumin is increased by bradykinin and inhibited by prostaglandins

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.2001.280.4.l732 ◽

2001 ◽

Vol 280 (4) ◽

pp. L732-L738 ◽

Cited By ~ 31

Author(s):

Pierre J. Farmer ◽

Sylvie G. Bernier ◽

Andrée Lepage ◽

Gaétan Guillemette ◽

Domenico Regoli ◽

...

Keyword(s):

Endothelial Cells ◽

Adenylate Cyclase ◽

Inhibitory Effect ◽

Dependent Manner ◽

Intracellular Level ◽

Concentration Dependent Manner ◽

Aortic Endothelial Cells ◽

Endothelial Monolayers ◽

Bovine Aortic Endothelial Cells

Using monolayers of bovine aortic endothelial cells (BAEC) in modified Boyden chambers, we examined the role of prostaglandins (PGs) in the bradykinin (BK)-induced increase of albumin permeability. BK induced a concentration-dependent increase of the permeability of BAEC, which reached 49.9 ± 1% at the concentration of 10−8 M. Two inhibitors of the prostaglandin G/H synthase, indomethacin (2.88 μM) and ibuprofen (10 μM), potentiated BK-induced permeability 1.8- and 3.9-fold, respectively. Exogenously administered PGE2and iloprost, a stable analog of prostacyclin, attenuated the effect of BK in a concentration-dependent manner. Butaprost equally reduced the effect of BK, suggesting the participation of the EP2receptor in this phenomenon. However, the EP4-selective antagonist AH-23848 did not significantly inhibit the protective effect of PGE2. The inhibitory effect of PGE2 was reversed by the adenylate cyclase inhibitor MDL-12330A (10 μM). These results suggest that BK-induced increase of permeability of BAEC monolayer to 125I-labeled albumin is negatively regulated by PGs. This postulated autocrine activity of PGs may involve an increase in the intracellular level of cAMP.

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

The International Journal of Artificial Organs ◽

10.1177/039139889702000704 ◽

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.

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

10.21203/rs.3.rs-22968/v1 ◽

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.

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

Biochemistry and Cell Biology ◽

10.1139/o92-122 ◽

1992 ◽

Vol 70 (9) ◽

pp. 803-809 ◽

Cited By ~ 17

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.

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Inhibitory Effect of Lidocaine on Cultured Porcine Aortic Endothelial Cell-dependent Antiaggregation of Platelets

Anesthesiology ◽

10.1097/00000542-199508000-00018 ◽

1995 ◽

Vol 83 (2) ◽

pp. 374-381. ◽

Cited By ~ 15

Author(s):

Toshiharu Az-ma ◽

MD Hardian ◽

Osafumi Yuge

Keyword(s):

Endothelial Cells ◽

Platelet Rich Plasma ◽

Inhibitory Effect ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Aortic Endothelial Cells ◽

Topical Lidocaine ◽

Vascular Spasm ◽

Stimulation Of ◽

Aortic Endothelial

Background Vascular spasm is a well-known complication during vascular surgery. Topical lidocaine is frequently used to prevent this spasm. However, the effects of lidocaine on the endothelium-dependent antiaggregation are not clear. Methods The aggregation of platelet-rich plasma (PRP) obtained from healthy volunteers was measured by the turbidimetric technique at 37 degrees C. (1) Cultured porcine aortic endothelial cells were preincubated with lidocaine (3.7 microM to 37 mM), NG-methyl-L-arginine (300 microM), or indomethacin (10 microM) for 30 min. The preincubation medium was exchanged with a medium containing +/- 1 microM bradykinin for 1-min stimulation of endothelial cells. One hundred microliters of the supernatant was then added to PRP (750 micro1) just after stimulation of PRP with collagen (4 micrograms/ml). (2) Authentic nitric oxide (NO) or prostacyclin (PGI2) was applied to collagen-stimulated PRP with or without lidocaine (100 micrograms/ml). Results (1) The supernatant from endothelial cells without bradykinin stimulation showed "basal" antiaggregation (13.8 +/- 3.2%; n = 6). Bradykinin enhanced the antiaggregation (100 +/- 0%; n = 6). NG-methyl-L-arginine or indomethacin (antagonists of NO or PGI2) inhibited the bradykinin-evoked antiaggregation (10.3 +/- 2.1% and 13.6 +/- 3.7%, respectively; n = 6). Simultaneous preincubation of both agents completely blocked the effect (-4.2 +/- 2.8%; n = 6). Lidocaine failed to influence basal antiaggregation, but it inhibited bradykinin-stimulated antiaggregation in a concentration-dependent manner (concentration causing 50% inhibition = 108 +/- 41 microM; n = 6). (2) In contrast, lidocaine did not shift the 50% effective concentration of NO (control, 1.3 +/- 0.1 microM vs. lidocaine, 1.6 +/- 0.1 microM) or PGI2 (control, 405 +/- 54 nM vs. lidocaine, 257 +/- 41 nM) for antiaggregation. Conclusions Our results suggest that lidocaine has an inhibitory effect on antiaggregation derived from endothelial cells, caused by the inhibition of NO and PGI2 released from endothelial cells.

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Modulation of calcium homeostasis in cultured rat aortic endothelial cells by intracellular acidification

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.1993.265.4.h1424 ◽

1993 ◽

Vol 265 (4) ◽

pp. H1424-H1433 ◽

Cited By ~ 2

Author(s):

R. C. Ziegelstein ◽

L. Cheng ◽

P. S. Blank ◽

H. A. Spurgeon ◽

E. G. Lakatta ◽

...

Keyword(s):

Endothelial Cells ◽

Vascular Endothelial Cells ◽

Cytosolic Calcium ◽

Ph Sensitive ◽

Vascular Endothelial ◽

Intracellular Acidification ◽

Cytosolic Ph ◽

Aortic Endothelial Cells ◽

Endothelial Monolayers ◽

Aortic Endothelial

Acidosis produces vasodilation in a process that may involve the vascular endothelium. Because synthesis and release of endothelium-derived vasodilatory substances are linked to an increase in cytosolic calcium concentration ([Ca2+]i), we examined the effect of intracellular acidification on cultured rat aortic endothelial cells loaded either with the pH-sensitive probe carboxy-seminaphthorhodafluor-1 or the Ca(2+)-sensitive fluorescent probe indo 1. The basal cytosolic pH (pHi) of endothelial monolayers in a 5% CO2-HCO3- buffer was 7.27 +/- 0.02 and that in a bicarbonate-free solution was 7.22 +/- 0.03. Acidification was induced either by removal of NH4Cl (delta pHi = -0.10 +/- 0.02), changing from a bicarbonate-free to a 5% CO2-HCO3(-)-buffered solution at constant buffer pH (delta pHi = -0.18 +/- 0.03), or changing from a 5% to a 20% CO2-HCO3- solution (delta pHi = -0.27 +/- 0.07). Regardless of the method used, intracellular acidification increased [Ca2+]i as indexed by indo 1 fluorescence. The increase in [Ca2+]i induced by changing from a 5 to a 20% CO2-HCO3- solution was not significantly altered by removal of buffer Ca2+ either before or after depletion of bradykinin- and thapsigargin-sensitive intracellular Ca2+ stores. Thus intracellular acidification of vascular endothelial cells releases Ca2+ into the cytosol either from pH-sensitive intracellular buffer sites, mitochondria, or from bradykinin- and thapsigargin-insensitive intracellular stores. This Ca2+ mobilization may be linked to endothelial synthesis and release of vasodilatory substances during acidosis.

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

Oxidative Medicine and Cellular Longevity ◽

10.1155/2020/2984613 ◽

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.

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Vascular Angiotensin-Converting Enzyme Activity in Man and other Species

Clinical Science ◽

10.1042/cs0660039 ◽

1984 ◽

Vol 66 (1) ◽

pp. 39-45 ◽

Cited By ~ 57

Author(s):

Mizuo Miyazaki ◽

Hideki Okunishi ◽

Kazuo Nishimura ◽

Noboru Toda

Keyword(s):

Angiotensin Converting Enzyme ◽

High Performance ◽

Muscle Tone ◽

Chloride Ion ◽

Kidney Cortex ◽

Dependent Manner ◽

Converting Enzyme ◽

Aortic Endothelial Cells ◽

Ace Activity ◽

Dose Dependent

1. Angiotensin-converting enzyme (ACE) activity in blood vessels of different species was determined. 2. ACE was solubilized by Nonidet P-40, and assayed by reversible phase high performance liquid chromatography. Approximately 98% ACE was recovered in the liquid phase by the use of the detergent. 3. The ACE activity varied with chloride ion (Cl−) concentrations; the maximum activities in dog, human, monkey and rabbit tissues were obtained at the concentrations of 800, 600, 600 and 300 mmol/l respectively. The optimal Cl− concentration was quite similar in different tissues and plasma obtained from the same species. 4. The ACE activity in the cerebral, mesenteric, pulmonary and renal arteries was in a range between 1.01 and 1.60 m-units/mg of protein in dogs and between 0.43 and 0.94 m-unit/mg of protein in monkeys. The activity in dog aortae was 0.20 ± 0.02 m-unit/mg of protein, and the activity in aortic endothelial cells was 2.61 ± 0.65 m-units/mg of protein. ACE activities in the dog lung, kidney cortex and cerebral cortex were 28.6 ± 2.6, 15.7 ± 3.0 and 3.5 ± 0.6m-units/mg of protein respectively. SA-446, a captopril-like ACE inhibitor, reduced the ACE activity in arteries in a dose-dependent manner. 5. Vascular ACE appears to be concentrated in the endothelium and may contribute to regulate vascular muscle tone and local blood flow by a conversion of angiotensin I into II.

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

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2021/6610116 ◽

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.

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