scholarly journals Saccharide expression on wounded endothelial cell monolayers in vitro

1989 ◽  
Vol 93 (1) ◽  
pp. 163-172
Author(s):  
R.Y. Ball ◽  
R.W. Stoddart ◽  
C.J. Jones ◽  
M.J. Mitchinson
Keyword(s):  
Cell Proliferation ◽  
Endothelial Cells ◽  
Cellular Response ◽  
Cell Spreading ◽  
Aortic Endothelial Cells ◽  
Cell Monolayers ◽  
Sequence Of Events ◽  
Monolayer Cultures ◽  
Porcine Aortic Endothelial Cells

Monolayer cultures of porcine aortic endothelial cells were used as a model of the endothelium of large arteries. Linear wounds were produced in such cultures by scraping and the subsequent sequence of events in nearby cells was analysed. The earliest detectable event was cellular spreading at the margins of the wounds (2 h) followed by cell migration (starting at 6–8 h) and cell proliferation in regions adjacent to the wound (16 h and later). Cell spreading was associated with the appearance of saccharides selectively at the spreading margins of the cells, which bound the lectins, ConA, LCA and PSA, and were sensitive to alpha-mannosidase. Terminal alpha-mannosyl residues were therefore present. The appearance of these saccharides suggests a mechanism by which monocytes might adhere to and/or migrate through the endothelium of vessels at sites of cellular response to injury.

scholarly journals Atorvastatin Down-Regulates the Primate Cellular Response to Porcine Aortic Endothelial Cells In Vitro

2008 ◽  
Vol 86 (5) ◽  
pp. 733-737 ◽  
Author(s):  
Mohamed Ezzelarab ◽  
Daniel Welchons ◽  
Corine Torres ◽  
Hidetaka Hara ◽  
Cassandra Long ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cellular Response ◽  
Aortic Endothelial Cells ◽  
Porcine Aortic Endothelial Cells ◽  
Aortic Endothelial

Disintegration of cytoskeletal structure of actin filaments in energy-depleted endothelial cells

1993 ◽  
Vol 264 (5) ◽  
pp. H1599-H1608 ◽  
Author(s):  
W. Kuhne ◽  
M. Besselmann ◽  
T. Noll ◽  
A. Muhs ◽  
H. Watanabe ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Actin Filaments ◽  
Fold Increase ◽  
Metabolic Inhibition ◽  
Ionophore A23187 ◽  
Aortic Endothelial Cells ◽  
Monolayer Cultures ◽  
Cell Retraction ◽  
Porcine Aortic Endothelial Cells ◽  
Reversible Phase

In a previous study [Watanabe, H., W. Kuhne, R. Spahr, P. Schwartz, and H. M. Piper. Am. J. Physiol. 260 (Heart Circ. Physiol. 29): H1344-H1352, 1991] metabolic inhibition (5 mM KCN + 5 mM 2-deoxy-D-glucose, for 2 h) was found to cause disintegration of F-actin filaments, cell retraction, and augmented paracellular macromolecule permeability in monolayer cultures of porcine aortic endothelial cells after a rapid depletion of ATP stores (90% in 5 min). These changes were reversible. In the present study, the nature of this cytoskeletal disintegration was investigated. 1) Disintegration of F-actin filaments within 2-h incubation under metabolic inhibition was accompanied by appearance of F-actin clumps in the cells, but total contents of F-actin remained unaltered. 2) Cytosolic Ca2+ levels rapidly rose in metabolically inhibited cells; after 2 h a 10-fold increase was observed. 3) Presence of the Ca2+ ionophore A23187 (10 microM) mimicked the reversible effect of metabolic inhibition on F-actin filaments and monolayer permeability but not the extensive depletion of ATP stores. 4) Existence of the Ca(2+)-activatable actin-severing protein gelsolin in endothelial cells was demonstrated. The results show that during the reversible phase of endothelial energy depletion disintegration of F-actin filaments is only partial, since it is based on their fragmentation and not depolymerization. Increase in cytosolic Ca2+ levels seems to be the primary cause for the fragmentation, possibly through the activation of gelsolin.

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 African Swine Fever Virus Infection of Porcine Aortic Endothelial Cells Leads to Inhibition of Inflammatory Responses, Activation of the Thrombotic State, and Apoptosis

2001 ◽  
Vol 75 (21) ◽  
pp. 10372-10382 ◽  
Author(s):  
Isabelle Vallée ◽  
Stephen W. G. Tait ◽  
Penelope P. Powell
Keyword(s):  
Endothelial Cells ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Surface Expression ◽  
Fever Virus ◽  
Parp Cleavage ◽  
Aortic Endothelial Cells ◽  
Porcine Aortic Endothelial Cells ◽  
Aortic Endothelial

ABSTRACT African swine fever (ASF) is an asymptomatic infection of warthogs and bushpigs, which has become an emergent disease of domestic pigs, characterized by hemorrhage, lymphopenia, and disseminated intravascular coagulation. It is caused by a large icosohedral double-stranded DNA virus, African swine fever virus (ASFV), with infection of macrophages well characterized in vitro and in vivo. This study shows that virulent isolates of ASFV also infect primary cultures of porcine aortic endothelial cells and bushpig endothelial cells (BPECs) in vitro. Kinetics of early and late gene expression, viral factory formation, replication, and secretion were similar in endothelial cells and macrophages. However, ASFV-infected endothelial cells died by apoptosis, detected morphologically by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling and nuclear condensation and biochemically by poly(ADP-ribose) polymerase (PARP) cleavage at 4 h postinfection (hpi). Immediate-early proinflammatory responses were inhibited, characterized by a lack of E-selectin surface expression and interleukin 6 (IL-6) and IL-8 mRNA synthesis. Moreover, ASFV actively downregulated interferon-induced major histocompatibility complex class I surface expression, a strategy by which viruses evade the immune system. Significantly, Western blot analysis showed that the 65-kDa subunit of the transcription factor NF-κB, a central regulator of the early response to viral infection, decreased by 8 hpi and disappeared by 18 hpi. Both disappearance of NF-κB p65 and cleavage of PARP were reversed by the caspase inhibitor z-VAD-fmk. Interestingly, surface expression and mRNA transcription of tissue factor, an important initiator of the coagulation cascade, increased 4 h after ASFV infection. These data suggest a central role for vascular endothelial cells in the hemorrhagic pathogenesis of the disease. Since BPECs infected with ASFV also undergo apoptosis, resistance of the natural host must involve complex pathological factors other than viral tropism.

Observations on Biological Effects of Carbon-13, Based on the Effects of 13C-Testosterone on Growth of Human Osteoblasts, Human Aortic Endothelial Cells, and Human Umbilical Vein Endothelial Cells in Vitro

2021 ◽  
Author(s):  
Mengqi Zhang ◽  
Wenning Yang ◽  
Xinchen Wu ◽  
Tengfei Zhang
Keyword(s):  
Cell Proliferation ◽  
Endothelial Cells ◽  
Isotope Effect ◽  
Umbilical Vein ◽  
Biological Effects ◽  
Bioactive Compound ◽  
Human Osteoblasts ◽  
Aortic Endothelial Cells ◽  
Umbilical Vein Endothelial Cells

Abstract Despite the increasing knowledge of biological isotope effect, comprehensive understanding of heavy isotope effect in the biological contexts has remained far less than expectation. The present study investigated the carbon isotope effect of 13C enriched testosterone on human cells. It was among the rare studies on carbon isotope effect of bioactive compound. Human osteoblasts, human aortic endothelial cells, and human umbilical vein endothelial cells were cultured in vitro and treated with testosterone and 13C enriched testosterone (13C/12C:6.7%). The impacts of physiological to pharmacological concentrations (10-10-10-5mol/L) of the bioactive compound were taken into account. The cell proliferation activities were measured using MTS assay. The levels of alkaline phosphatase and osteocalcin in osteoblasts were tested. Our results established that 13C enriched testosterone exhibited different biological effects from testosterone. At the concentrations of 10-10mol/L and 10-5mol/L, there were significant differences in prompting cell proliferation between testosterone and 13C enriched testosterone. At physiological concentrations, testosterone prompted proliferations of the three kinds of cells; whereas, 13C enriched testosterone did not prompt the cell proliferation, and its effects were not concentration dependent. At supraphysiological concentration (10-5mol/L), testosterone had the trend of inhibiting cell growth; whereas, 13C enriched testosterone had the trend of prompting cell growth. 13C enriched testosterone significantly enhanced osteocalcin secretion in human osteoblasts at supraphysiological concentration. These findings challenged the common view of growth retardation effect of heavy isotope, which imply that biological isotope effects are worthy of further study. The potential applications of 13C enriched compound were discussed.

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