25-Hydroxycholesterol exerts both a cox-2-dependent transient proliferative effect and cox-2-independent cytotoxic effect on bovine endothelial cells in a time- and cell-type-dependent manner

The exposure of human endothelial cells to 3-morpholinosydnonimine (SIN-1) induced the expression of cyclooxygenase-2 (COX-2) in a dose- and time-dependent manner. Interestingly, after a prolonged incubation (>8 h) several proteoforms were visualized by Western blot, corresponding to different states of glycosylation of the protein. This effect was specific for SIN-1 that generates peroxynitrite and it was not detected with other nitric oxide-donors. Metabolic labeling experiments using 35S or cycloheximide suggested that the formation of hypoglycosylated COX-2 was dependent on de novo synthesis of the protein rather than the deglycosylation of the native protein. Moreover, SIN-1 reduced the activity of the hexokinase, the enzyme responsible for the first step of glycolysis. The hypoglycosylated COX-2 induced by SIN-1 showed a reduced capacity to generate prostaglandins and the activity was only partially recovered after immunoprecipitation. Finally, hypoglycosylated COX-2 showed a more rapid rate of degradation compared to COX-2 induced by IL-1α and an alteration in the localization with an accumulation mainly detected in the nuclear membrane. Our results have important implication to understand the effect of peroxynitrite on COX-2 expression and activity, and they may help to identify new pharmacological tools direct to increase COX-2 degradation or to inhibit its activity.

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Intracisternal administration of transforming growth factor-β evokes fever through the induction of cyclooxygenase-2 in brain endothelial cells

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00181.2007 ◽

2008 ◽

Vol 294 (1) ◽

pp. R266-R275 ◽

Cited By ~ 3

Author(s):

Shigenobu Matsumura ◽

Tetsuro Shibakusa ◽

Teppei Fujikawa ◽

Hiroyuki Yamada ◽

Kiyoshi Matsumura ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Growth Factor ◽

Proinflammatory Cytokines ◽

Transforming Growth Factor ◽

Transforming Growth Factor Β ◽

Cyclooxygenase 2 ◽

Dependent Manner ◽

Cox 2 ◽

Β Receptor

Transforming growth factor-β (TGF-β), a pleiotropic cytokine, regulates cell proliferation, differentiation, and apoptosis, and plays a key role in development and tissue homeostasis. TGF-β functions as an anti-inflammatory cytokine because it suppresses microglia and B-lymphocyte functions, as well as the production of proinflammatory cytokines. However, we previously demonstrated that the intracisternal administration of TGF-β induces fever like that produced by proinflammatory cytokines. In this study, we investigated the mechanism of TGF-β-induced fever. The intracisternal administration of TGF-β increased body temperature in a dose-dependent manner. Pretreatment with cyclooxygenase-2 (COX-2)-selective inhibitor significantly suppressed TGF-β-induced fever. COX-2 is known as one of the rate-limiting enzymes of the PGE2 synthesis pathway, suggesting that fever induced by TGF-β is COX-2 and PGE2 dependent. TGF-β increased PGE2 levels in cerebrospinal fluid and increased the expression of COX-2 in the brain. Double immunostaining of COX-2 and von Willebrand factor (vWF, an endothelial cell marker) revealed that COX-2-expressing cells were mainly endothelial cells. Although not all COX-2-immunoreactive cells express TGF-β receptor, some COX-2-immunoreactive cells express activin receptor-like kinase-1 (ALK-1, an endothelial cell-specific TGF-β receptor), suggesting that TGF-β directly or indirectly acts on endothelial cells to induce COX-2 expression. These findings suggest a novel function of TGF-β as a proinflammatory cytokine in the central nervous system.

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Haemophilus somnus Induces Apoptosis in Bovine Endothelial Cells In Vitro

Infection and Immunity ◽

10.1128/iai.69.3.1650-1660.2001 ◽

2001 ◽

Vol 69 (3) ◽

pp. 1650-1660 ◽

Cited By ~ 46

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.

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The Induction of Cyclooxygenase-2 in IL-1β-Treated Endothelial Cells is Inhibited by Prostaglandin E2 through cAMP

Mediators of Inflammation ◽

10.1080/09629359990298 ◽

1999 ◽

Vol 8 (6) ◽

pp. 287-294 ◽

Cited By ~ 32

Author(s):

Pravit Akarasereenont ◽

Kitirat Techatrisak ◽

Sirikul Chotewuttakorn ◽

Athiwat Thaworn

Keyword(s):

Endothelial Cells ◽

Biological Activities ◽

Umbilical Vein ◽

The Body ◽

Prostaglandin E ◽

Dependent Manner ◽

Negative Feedback Regulation ◽

Cox 2 ◽

Arachidonic Acids ◽

Cox 1

Prostaglandins (PGS) have numerous cardiovascular and inflammatory effects. Cyclooxygenase (COX), which exists as COX-1 and COX-2 isoforms, is the first enzyme in the pathway in which arachidonic acid is converted to PGs. Prostaglandin E2 (PGE2) exerts a variety of biological activities for the maintenance of local homeostasis in the body. Elucidation of PGE2 involvement in the signalling molecules such as COX could lead to potential therapeutic interventions. Here, we have investigated the effects of PGE2 on the induction of COX-2 in human umbilical vein endothelial cells (HUVEC) treated with interleukin-1β (IL-1β 1 ng/ml). COX activity was measured by the production of 6-keto-PGF1α, PGE2, PGF2α and thromboxane B2 (TXB2) in the presence of exogenous arachidonic acids (10 μM for 10 min) using enzyme immunoassay (EIA). COX-1 and COX-2 protein was measured by immunoblotting using specific antibody. Untreated HUVEC contained only COX-1 protein while IL-1β treated HUVEC contained COX-1 and COX-2 protein. PGE2 (3 μM for 24 h) did not affect on COX activity and protein in untreated HUVEC. Interestingly, PGE2 (3 μM for 24 h) can inhibit COX-2 protein, but not COX-1 protein, expressed in HUVEC treated with IL1 β. This inhibition was reversed by coincubation with forskolin (100 μM). The increased COX activity in HUVEC treated with IL-1β was also inhibited by PGE2 (0.03, 0.3 and 3 μM for 24 h) in a dose-dependent manner. Similarly, forskolin (10, 50 or 100 μM) can also reverse the inhibition of PGE2 on increased COX activity in IL-1β treated HUVEC. The results suggested that (i) PGE2 can initiate negative feedback regulation in the induction of COX-2 elicited by IL-1β in endothelial cells, (ii) the inhibition of PGE2 on COX-2 protein and activity in IL-1β treated HUVEC is mediated by cAMP and (iii) the therapeutic use of PGE2 in the condition which COX-2 has been involved may have different roles.

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Rickettsia rickettsii Infection of Human Macrovascular and Microvascular Endothelial Cells Reveals Activation of Both Common and Cell Type-Specific Host Response Mechanisms

Infection and Immunity ◽

10.1128/iai.01335-09 ◽

2010 ◽

Vol 78 (6) ◽

pp. 2599-2606 ◽

Cited By ~ 29

Author(s):

Elena Rydkina ◽

Loel C. Turpin ◽

Sanjeev K. Sahni

Keyword(s):

Endothelial Cells ◽

Host Response ◽

Cell Types ◽

Microvascular Endothelial Cells ◽

Cell Type ◽

Rickettsia Rickettsii ◽

Specific Host ◽

Cell Type Specific ◽

Cox 2 ◽

Microvascular Endothelial

ABSTRACT Although inflammation and altered barrier functions of the vasculature, due predominantly to the infection of endothelial cell lining of small and medium-sized blood vessels, represent salient pathological features of human rickettsioses, the interactions between pathogenic rickettsiae and microvascular endothelial cells remain poorly understood. We have investigated the activation of nuclear transcription factor-kappa B (NF-κB) and p38 mitogen-activated protein (MAP) kinase, expression of heme oxygenase 1 (HO-1) and cyclooxygenase 2 (COX-2), and secretion of chemokines and prostaglandins after Rickettsia rickettsii infection of human cerebral, dermal, and pulmonary microvascular endothelial cells in comparison with pulmonary artery cells of macrovascular origin. NF-κB and p38 kinase activation and increased HO-1 mRNA expression were clearly evident in all cell types, along with relatively similar susceptibility to R. rickettsii infection in vitro but considerable variations in the intensities/kinetics of the aforementioned host responses. As expected, the overall activation profiles of macrovascular endothelial cells derived from human pulmonary artery and umbilical vein were nearly identical. Interestingly, cerebral endothelial cells displayed a marked refractoriness in chemokine production and secretion, while all other cell types secreted various levels of interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1) in response to infection. A unique feature of all microvascular endothelial cells was the lack of induced COX-2 expression and resultant inability to secrete prostaglandin E2 after R. rickettsii infection. Comparative evaluation thus yields the first experimental evidence for the activation of both common and unique cell type-specific host response mechanisms in macrovascular and microvascular endothelial cells infected with R. rickettsii, a prototypical species known to cause Rocky Mountain spotted fever in humans.

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The influence of selective COX-2 inhibitor on phase of healing surgical wounds: proliferation and secretion of bFGF by endothelial cells

Annales Universitatis Mariae Curie-Sklodowska sectio C – Biologia ◽

10.17951/c.2017.72.1.7-13 ◽

2018 ◽

Vol 72 (1) ◽

pp. 7

Author(s):

Łukasz Jasiak ◽

Mateusz Kowalczyk ◽

Paula Mazan ◽

Edward Kowalczyk ◽

Monika Sienkiewicz ◽

...

Keyword(s):

Wound Healing ◽

Endothelial Cells ◽

Cell Viability ◽

Vascular Endothelial Cells ◽

Vascular Endothelial ◽

Antiinflammatory Drugs ◽

Antiangiogenic Effect ◽

Proliferative Effect ◽

Mtt Method ◽

Cox 2

<p>The process of wound healing consists of the following phases: inflammation, proliferation, remodeling. Non-steroidal antiinflammatory drugs may be important in this process, especially in a stage called angiogenesis. For this reason, it was decided to investigate the effect of selective COX-2 (cyclooxygenase 2) inhibitor (NS-398) on the proliferation of endothelial cells and their ability to secrete bFGF (fibroblast growth factor) for vascular endothelial cells (HMEC-1). For determination of the secretion of bFGF in a cell line HMEC-1 immunosorbent ELISA assays were used. In turn, the cell proliferation assay was performed using the MTT method. Using MTT method, it was found that NS-398 at 10 μM did not affect cell viability. Whereas selective COX-2 inhibitor at 100 μM decreased cell viability in a statistically significant manner and inhibited the proliferative effect of 100 μg/mL LPS at concentrations of 10 and 100 μM. In the further step, application of NS-398 (10 and 100 μM) with LPS (100 μg/mL; inflammatory environment) reduced the secretion of bFGF in a statistically significant manner. The investigations showed that NS-398 has an antiangiogenic effect which is based on reducing the proliferation of vascular endothelial cells and inhibiting the secretion of bFGF- factor responsible for angiogenesis during wound healing.</p>

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Profiling of Primary and Mature miRNA Expression in Atherosclerosis Associated Cell Types

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvbaha.121.315579 ◽

2021 ◽

Author(s):

Pierre R. Moreau ◽

Vanesa Tomas Bosch ◽

Maria Bouvy-Liivrand ◽

Kadri Õunap ◽

Tiit Örd ◽

...

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Molecular Mechanisms ◽

Umbilical Vein ◽

Cell Types ◽

Integrative Approach ◽

Dependent Manner ◽

Cell Type ◽

A Cell ◽

Cell Type Specific

Objective: Atherosclerosis is the underlying cause of most cardiovascular diseases. The main cell types associated with disease progression in the vascular wall are endothelial cells, smooth muscle cells, and macrophages. Although their role in atherogenesis has been extensively described, molecular mechanisms underlying gene expression changes remain unknown. The objective of this study was to characterize microRNA (miRNA)-related regulatory mechanisms taking place in the aorta during atherosclerosis: Approach and Results: We analyzed the changes in primary human aortic endothelial cells and human umbilical vein endothelial cell, human aortic smooth muscle cell, and macrophages (CD14+) under various proatherogenic stimuli by integrating GRO-seq, miRNA-seq, and RNA-seq data. Despite the highly cell-type-specific expression of multi-variant pri-miRNAs, the majority of mature miRNAs were found to be common to all cell types and dominated by 2 to 5 abundant miRNA species. We demonstrate that transcription contributes significantly to the mature miRNA levels although this is dependent on miRNA stability. An analysis of miRNA effects in relation to target mRNA pools highlighted pathways and targets through which miRNAs could affect atherogenesis in a cell-type-dependent manner. Finally, we validate miR-100-5p as a cell-type specific regulator of inflammatory and HIPPO-YAP/TAZ-pathways. Conclusions: This integrative approach allowed us to characterize miRNA dynamics in response to a proatherogenic stimulus and identify potential mechanisms by which miRNAs affect atherogenesis in a cell-type-specific manner.

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Human apolipoprotein A-I induces cyclooxygenase-2 expression and prostaglandin I-2 release in endothelial cells through ATP-binding cassette transporter A1

AJP Cell Physiology ◽

10.1152/ajpcell.00055.2011 ◽

2011 ◽

Vol 301 (3) ◽

pp. C739-C748 ◽

Cited By ~ 34

Author(s):

Donghui Liu ◽

Liang Ji ◽

Xunliang Tong ◽

Bing Pan ◽

Jing-Yan Han ◽

...

Keyword(s):

Endothelial Cells ◽

Umbilical Vein ◽

Cyclooxygenase 2 ◽

Mitogen Activated Protein Kinase ◽

Atp Binding ◽

Dependent Manner ◽

Atp Binding Cassette Transporter ◽

Apolipoprotein A ◽

Cox 2 ◽

Atp Binding Cassette

High-density lipoprotein (HDL) can induce cyclooxygenase-2 (COX-2) expression and prostacyclin I-2 (PGI-2) release in endothelial cells to exert multiple antiatherogenic functions. This effect has been attributed mainly to the role of sphingosine-1-phosphate (S1P) integrated in HDL. However, whether apolipoprotein A-I (apoA-I), the major apolipoprotein of HDL, could induce COX-2 expression and PGI-2 release still remains unclear. In the present study, we selectively delipidated HDL and confirmed that apoA-I could facilitate COX-2 expression and PGI-2 production in human umbilical vein endothelial cells (HUVECs). ApoA-I, but not trypsinized apoA-I, induced COX-2 expression in a time- and dose-dependent manner consistent with a key role for apoA-I in this process. Additionally, cotreatment of apoA-I with S1P further enhanced COX-2 expression and PGI-2 production in HUVECs. These effects triggered by apoA-I were not inhibited by pertussis toxin, consistent with SIP receptor independent pathway for apoA-I effect. Moreover, we demonstrated that the activation of p38 mitogen-activated protein kinase (MAPK), extracellular receptor kinase (ERK) 1/2, and JAK2 pathways by apoA-I was involved in the expression of COX-2 and the release of PGI-2 in HUVECs, and these effects were inhibited by their specific inhibitors, respectively. Small interfering RNA experiments showed that ATP binding-cassette transporter A1 (ABCA1) was required for COX-2 expression and PGI-2 release induced by apoA-I. Thus our results indicate that apoA-I induces COX-2 expression and PGI-2 release through ABCA1 and the activation of intracellular p38 MAPK, ERK1/2, as well as JAK2 pathways, and apoA-I can reinforce these effects with S1P in HUVECs. These novel effects of apoA-I could in part mediate antiatherogenic effects of HDL.

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Proliferative and cytotoxic effects of mildly oxidized low-density lipoproteins on vascular smooth-muscle cells

Biochemical Journal ◽

10.1042/bj3091015 ◽

1995 ◽

Vol 309 (3) ◽

pp. 1015-1020 ◽

Cited By ~ 65

Author(s):

N Augé ◽

M T Pieraggi ◽

J C Thiers ◽

A Nègre-Salvayre ◽

R Salvayre

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Smooth Muscle Cells ◽

Cytotoxic Effect ◽

Density Lipoprotein ◽

Muscle Cells ◽

Butylated Hydroxytoluene ◽

Ldl Oxidation ◽

Low Density ◽

Proliferative Effect

We have investigated the role of low-density lipoprotein (LDL) oxidation in the proliferative effect of LDLs on cultured bovine aortic smooth-muscle cells and compared it with their effect on bovine aortic endothelial cells. The following conclusions were reached. (1) Non-toxic doses of mildly oxidized LDLs elicit a proliferative effect on smooth-muscle cells significantly higher than that of native LDLs or lipoprotein-depleted serum. The proliferative effect is dependent on time (relatively slow), dose (high doses are cytotoxic) and the level of LDL oxidation. (2) The proliferative effect on smooth-muscle cells is counterbalanced at high concentrations of mildly oxidized LDLs (or at high oxidation levels) by their cytotoxic effect. (3) The same dose of mildly oxidized LDLs exhibits no proliferative effect on endothelial cells but rather a cytotoxic one. Endothelial cells may therefore be intrinsically more susceptible to the cytotoxic effect of mildly oxidized LDLs than are smooth-muscle cells. (4) The proliferative effect of native LDLs on smooth-muscle cells results (at least in part) from cell-induced LDL oxidation during cell culture as suggested by (i) the progressive LDL oxidation over the 3 days of contact between LDLs and smooth-muscle cells and (ii) the concomitant inhibition of LDL oxidation and proliferative effect by butylated hydroxytoluene. The hypothetical mechanisms and potential involvement in atherogenesis are discussed.

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Methylglyoxal mediates vascular inflammation via JNK and p38 in human endothelial cells

AJP Cell Physiology ◽

10.1152/ajpcell.00252.2008 ◽

2008 ◽

Vol 295 (6) ◽

pp. C1510-C1517 ◽

Cited By ~ 74

Author(s):

Hideyuki Yamawaki ◽

Kazuaki Saito ◽

Muneyoshi Okada ◽

Yukio Hara

Keyword(s):

Endothelial Cells ◽

Inflammatory Responses ◽

Morphological Changes ◽

Cell Damage ◽

Umbilical Vein ◽

P38 Map Kinase ◽

Diabetic Patients ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Cox 2

Methylglyoxal (MGO) is a reactive metabolite of glucose. Since the plasma concentration of MGO is increased in diabetic patients, MGO is implicated in diabetes-associated vascular endothelial cells (ECs) injury, which might be responsible for atherosclerosis. In the present study, we examined effects of treatment of human umbilical vein ECs with MGO on EC morphology and inflammatory responses. MGO (24 h) induced cytotoxic morphological changes in a concentration-dependent manner (0–420 μM). MGO induced mRNA and protein expression of cyclooxygenase (COX)-2 in a concentration (0–420 μM)- and time (6–24 h)-dependent manner. COX-2 induction was associated with increased PGE2 release. Acute treatment with MGO (20 min) induced concentration-dependent (0–420 μM) activation of JNK and p38 MAP kinase but not ERK or NF-κB. Both the JNK inhibitor SP600125 and the p38 inhibitor SB203580 prevented the MGO induction of COX-2. However, inhibiting JNK and p38 or COX-2 was ineffective to the morphological damage by MGO (420 μM, 24 h). EUK134, a synthetic combined superoxide dismutase/catalase mimetic, had no effect on MGO-induced COX-2. Present results indicated that MGO mediates JNK- and p38-dependent EC inflammatory responses, which might be independent of oxidative stress. On the other hand, MGO-induced morphological cell damage seems unlikely to be associated with COX-2-PGE2.

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