p38 Mitogen-Activated Protein Kinase Mediates Palmitate-Induced Apoptosis But Not Inhibitor of Nuclear Factor-κB Degradation in Human Coronary Artery Endothelial Cells

Plasma free fatty acids are elevated in patients with type 2 diabetes and contribute to the pathogenesis of insulin resistance and endothelial dysfunction. The p38 MAPK mediates stress, inflammation, and apoptosis. Whether free fatty acids induce apoptosis and/or activate nuclear factor-κB inflammatory pathway in human coronary artery endothelial cells (hCAECs) and, if so, whether this involves the p38 MAPK pathway is unknown. hCAECs (passages 4–6) were grown to 70% confluence and then incubated with palmitate at concentrations of 0–300 μm for 6–48 h. Palmitate at 100, 200, or 300 μm markedly increased apoptosis after 12 h of incubation. This apoptotic effect was time (P = 0.008) and dose (P = 0.006) dependent. Palmitate (100 μm for 24 h) induced a greater than 2-fold increase in apoptosis, which was accompanied with a 4-fold increase in p38 MAPK activity (P < 0.001). Palmitate did not affect the phosphorylation of Akt1 or ERK1/2. SB203580 (a specific inhibitor of p38 MAPK) alone did not affect cellular apoptosis; however, it abolished palmitate-induced apoptosis and p38 MAPK activation. Palmitate significantly reduced the level of inhibitor of nuclear factor-κB (IκB). However, treatment of cells with SB203580 did not restore IκB to baseline. We conclude that palmitate induces hCAEC apoptosis via a p38 MAPK-dependent mechanism and may participate in coronary endothelial injury in diabetes. However, palmitate-mediated IκB degradation in hCAECs is independent of p38 MAPK activity.

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Saturated, but Not Unsaturated, Fatty Acids Induce Apoptosis of Human Coronary Artery Endothelial Cells via Nuclear Factor- B Activation

Diabetes ◽

10.2337/db06-0188 ◽

2006 ◽

Vol 55 (11) ◽

pp. 3121-3126 ◽

Cited By ~ 102

Author(s):

K. Staiger ◽

H. Staiger ◽

C. Weigert ◽

C. Haas ◽

H.-U. Haring ◽

...

Keyword(s):

Fatty Acids ◽

Endothelial Cells ◽

Coronary Artery ◽

Unsaturated Fatty Acids ◽

Nuclear Factor ◽

Human Coronary Artery ◽

Factor B

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γ-Tocopherol Decreases Ox-LDL-Mediated Activation of Nuclear Factor-κB and Apoptosis in Human Coronary Artery Endothelial Cells

Biochemical and Biophysical Research Communications ◽

10.1006/bbrc.1999.0716 ◽

1999 ◽

Vol 259 (1) ◽

pp. 157-161 ◽

Cited By ~ 49

Author(s):

Dayuan Li ◽

Tom Saldeen ◽

Jawahar L. Mehta

Keyword(s):

Endothelial Cells ◽

Coronary Artery ◽

Nuclear Factor Κb ◽

Nuclear Factor ◽

Human Coronary Artery

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Thrombin Stimulation of the Vascular Cell Adhesion Molecule-1 Promoter in Endothelial Cells Is Mediated by Tandem Nuclear Factor-κB and GATA Motifs

Journal of Biological Chemistry ◽

10.1074/jbc.m108363200 ◽

2001 ◽

Vol 276 (50) ◽

pp. 47632-47641 ◽

Cited By ~ 78

Author(s):

Takashi Minami ◽

William C. Aird

Keyword(s):

Endothelial Cells ◽

Adhesion Molecule ◽

Umbilical Vein ◽

Fold Increase ◽

Nuclear Factor Κb ◽

Nuclear Factor ◽

Mobility Shift ◽

Response Element ◽

Vascular Adhesion ◽

Stimulation Of

The goal of this study was to delineate the transcriptional mechanisms underlying thrombin-mediated induction of vascular adhesion molecule-1 (VCAM-1). Treatment of human umbilical vein endothelial cells with thrombin resulted in a 3.3-fold increase in VCAM-1 promoter activity. The upstream promoter region of VCAM-1 contains a thrombin response element, two nuclear factor κB (NF-κB) motifs, and a tandem GATA motif. In transient transfection assays, mutation of the thrombin response element had no effect on thrombin induction. In contrast, mutation of either NF-κB site resulted in a complete loss of induction, whereas a mutation of the two GATA motifs resulted in a significant reduction in thrombin stimulation. In electrophoretic mobility shift assays, nuclear extracts from thrombin-treated endothelial cells displayed markedly increased binding to the tandem NF-κB and GATA motifs. The NF-κB complex was supershifted with anti-p65 antibodies, but not with antibodies to RelB, c-Rel, p50, or p52. The GATA complex was supershifted with antibodies to GATA-2, but not GATA-3 or GATA-6. A construct containing tandem copies of the VCAM-1 GATA motifs linked to a minimal thymidine kinase promoter was induced 2.4-fold by thrombin. Taken together, these results suggest that thrombin stimulation of VCAM-1 in endothelial cells is mediated by the coordinate action of NF-κB and GATA transcription factors.

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Long intergenic noncoding RNA p21 mediates oxidized LDL-induced apoptosis and expression of LOX-1 in human coronary artery endothelial cells

Molecular Medicine Reports ◽

10.3892/mmr.2017.7623 ◽

2017 ◽

Vol 16 (6) ◽

pp. 8513-8519 ◽

Cited By ~ 5

Author(s):

Tao Zhou ◽

Xiaofang Chen

Keyword(s):

Endothelial Cells ◽

Coronary Artery ◽

Noncoding Rna ◽

Oxidized Ldl ◽

Human Coronary Artery ◽

Long Intergenic Noncoding Rna ◽

Induced Apoptosis

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Carvedilol prevents epinephrine-induced apoptosis in human coronary artery endothelial cells modulation of Fas/Fas ligand and caspase-3 pathway

Cardiovascular Research ◽

10.1016/s0008-6363(99)00369-7 ◽

2000 ◽

Vol 45 (3) ◽

pp. 788-794 ◽

Cited By ~ 46

Author(s):

F Romeo

Keyword(s):

Endothelial Cells ◽

Coronary Artery ◽

Caspase 3 ◽

Fas Ligand ◽

Human Coronary Artery ◽

Induced Apoptosis

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Lipoapoptosis of human coronary artery endothelial cells induced by saturated fatty acids is mediated by NFκB activation

Diabetologie und Stoffwechsel ◽

10.1055/s-2006-943974 ◽

2006 ◽

Vol 1 (S 1) ◽

Author(s):

K Staiger ◽

H Staiger ◽

C Weigert ◽

HU Häring ◽

M Kellerer

Keyword(s):

Fatty Acids ◽

Endothelial Cells ◽

Coronary Artery ◽

Saturated Fatty Acids ◽

Human Coronary Artery

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SPECIFIC TOXICITY OF CALCIUM PHOSPHATE BIONS FOR HUMAN VENOUS AND ARTERIAL ENDOTHELIAL CELLS

ZHurnal «Patologicheskaia fiziologiia i eksperimental`naia terapiia» ◽

10.25557/0031-2991.2019.01.53-61 ◽

2019 ◽

pp. 53-61

Author(s):

Д.К. Шишкова ◽

Е.А. Великанова ◽

В.Г. Матвеева ◽

Ю.А. Кудрявцева ◽

А.Г. Кутихин

Keyword(s):

Endothelial Cells ◽

Coronary Artery ◽

Calcium Phosphate ◽

Exposure Time ◽

Internal Thoracic Artery ◽

Human Coronary Artery ◽

Hoechst 33342 ◽

Ethidium Bromide Staining ◽

Induced Apoptosis ◽

Arterial Endothelial Cells

Цель исследования - оценка токсичности кальций-фосфатных бионов (КФБ) и магний-фосфатных бионов (МФБ) для культур эндотелиальных клеток. Методика. Эндотелиотоксичность бионов изучена при помощи добавления равных концентраций МФБ или КФБ к: 1) разреженным или конфлюэнтным культурам иммортализованных венозных эндотелиальных клеток человека линии EA.hy 926 с последующим культивированием в течение 24 ч или 4 ч соответственно; 2) конфлюэнтным культурам коммерческих первичных эндотелиальных клеток коронарной и внутренней грудной артерии человека с последующим культивированием в течение 24 ч. Эндотелиотоксические эффекты бионов оценивали при помощи сочетанного окрашивания клеток флюоресцентными красителями Hoechst 33342 и бромистым этидием, а также посредством колориметрического теста. Кроме того, методом проточной цитометрии оценивали пути и стадии гибели клеток вышеуказанных культур. Результаты. В отличие от МФБ, КФБ индуцировали гибель эндотелиальных клеток всех 3 линий путем апоптоза. Устойчивость культур к токсическому действию КФБ определялась степенью их конфлюэнтности (конфлюэнтные культуры более устойчивы чем разреженные) и типом клеточной линии (эндотелиальные клетки внутренней грудной артерии продемонстрировали большую устойчивость в сравнении с эндотелиальными клетками коронарной артерии). Заключение. Токсичность КФБ для культур эндотелиальных клеток специфична, то есть определяется их специфическим минеральным составом, а не общей для всех типов бионов корпускулярной природой. Добавление КФБ к конфлюэнтным культурам первичных артериальных эндотелиальных клеток и к иммортализованным венозным эндотелиальным клеткам вызывало их гибель, при этом экспозиция МФБ не оказывает значимого токсического действия. Эндотелиальные клетки внутренней грудной артерии менее чувствительны к воздействию КФБ в сравнении с эндотелиальными клетками коронарной артерии человека. Aim. To compare toxicity of calcium phosphate bions (CPB) and magnesium phosphate bions (MPB) for endothelial cells. Me-thods. To assess endothelial toxicity of the bions, we first added equal concentrations of either MPB or CPB to: 1) non-confluent or confluent cultures of immortalized human venous endothelial cells EA.hy 926 with the exposure time of 24 h or 4 h, respectively; 2) confluent cultures of commercially available primary human coronary artery and internal thoracic artery endothelial cells, with the exposure time of 24 h. Endothelial toxicity was then evaluated by combined Hoechst 33342 and ethidium bromide staining following fluorescence microscopy and by colorimetric cytotoxicity assay. In addition, we attempted to determine the pathway of bion-induced cell death utilizing flow cytometry. Results. In contrast to the MPB, CPB induced apoptosis of all studied endothelial cell lines. Resistance of endothelial cells to the CPB was defined by their confluence (confluent cultures demonstrated higher resistance), and cell type (internal thoracic artery endothelial cells were more resistant to the CPB as compared to the coronary artery endothelial cells). Conclusions. Endothelial toxicity of the CPB is defined by their specific mineral composition but not by their corpuscular nature, which is common for all nanoparticles. Addition of the CPB to the confluent cultures of primary human arterial cells and to immortalized human venous endothelial cells evoked their death. On the contrary, exposure to the MPB did not cause any toxic effects. Human internal thoracic artery endothelial cells are more resistant to the CPB in comparison with coronary artery endothelial cells.

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Activation of nuclear factor-κB during doxorubicin-induced apoptosis in endothelial cells and myocytes is pro-apoptotic: the role of hydrogen peroxide

Biochemical Journal ◽

10.1042/bj20020752 ◽

2002 ◽

Vol 367 (3) ◽

pp. 729-740 ◽

Cited By ~ 165

Author(s):

Suwei WANG ◽

Srigiridhar KOTAMRAJU ◽

Eugene KONOREV ◽

Shasi KALIVENDI ◽

Joy JOSEPH ◽

...

Keyword(s):

Endothelial Cells ◽

Nuclear Translocation ◽

Nuclear Factor Κb ◽

Nuclear Factor ◽

Apoptotic Pathway ◽

Toxic Side Effect ◽

Rat Cardiomyocytes ◽

A Cell ◽

Induced Apoptosis

Doxorubicin (DOX) is a widely used anti-tumour drug. Cardiotoxicity is a major toxic side effect of DOX therapy. Although recent studies implicated an apoptotic pathway in DOX-induced cardiotoxicity, the mechanism of DOX-induced apoptosis remains unclear. In the present study, we investigated the role of reactive oxygen species and the nuclear transcription factor nuclear factor κB (NF-κB) during apoptosis induced by DOX in bovine aortic endothelial cells (BAECs) and adult rat cardiomyocytes. DOX-induced NF-κB activation is both dose- and time-dependent, as demonstrated using electrophoretic mobility-shift assay and luciferase and p65 (Rel A) nuclear-translocation assays. Addition of a cell-permeant iron metalloporphyrin significantly suppressed NF-κB activation and apoptosis induced by DOX. Overexpression of glutathione peroxidase, which detoxifies cellular H2O2, significantly decreased DOX-induced NF-κB activation and apoptosis. Inhibition of DOX-induced NF-κB activation by a cell-permeant peptide SN50 that blocks translocation of the NF-κB complex into the nucleus greatly diminished DOX-induced apoptosis. Apoptosis was inhibited when IκB mutant vector, another NF-κB inhibitor, was added to DOX-treated BAECs. These results suggest that NF-κB activation in DOX-treated endothelial cells and myocytes is pro-apoptotic, in contrast with DOX-treated cancer cells, where NF-κB activation is anti-apoptotic. Removal of intracellular H2O2 protects endothelial cells and myocytes from DOX-induced apoptosis, possibly by inhibiting NF-κB activation. These findings suggest a novel mechanism for enhancing the therapeutic efficacy of DOX.

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Exendin-4 protects endothelial cells from lipoapoptosis by PKA, PI3K, eNOS, p38 MAPK, and JNK pathways

Journal of Molecular Endocrinology ◽

10.1530/jme-12-0166 ◽

2013 ◽

Vol 50 (2) ◽

pp. 229-241 ◽

Cited By ~ 42

Author(s):

Özlem Erdogdu ◽

Linnéa Eriksson ◽

Hua Xu ◽

Åke Sjöholm ◽

Qimin Zhang ◽

...

Keyword(s):

Endothelial Cells ◽

P38 Mapk ◽

Experimental Studies ◽

Receptor Activation ◽

No Production ◽

Ros Production ◽

Human Coronary Artery ◽

Glucagon Like Peptide 1 ◽

Underlying Mechanisms ◽

Induced Apoptosis

Experimental studies have indicated that endothelial cells play an important role in maintaining vascular homeostasis. We previously reported that human coronary artery endothelial cells (HCAECs) express the glucagon-like peptide 1 (GLP1) receptor and that the stable GLP1 mimetic exendin-4 is able to activate the receptor, leading to increased cell proliferation. Here, we have studied the effect of exendin-4 and native GLP1 (7–36) on lipoapoptosis and its underlying mechanisms in HCAECs. Apoptosis was assessed by DNA fragmentation and caspase-3 activation, after incubating cells with palmitate. Nitric oxide (NO) and reactive oxidative species (ROS) were analyzed. GLP1 receptor activation, PKA-, PI3K/Akt-, eNOS-, p38 MAPK-, and JNK-dependent pathways, and genetic silencing of transfection of eNOS were also studied. Palmitate-induced apoptosis stimulated cells to release NO and ROS, concomitant with upregulation of eNOS, which required activation of p38 MAPK and JNK. Exendin-4 restored the imbalance between NO and ROS production in which ROS production decreased and NO production was further augmented. Incubation with exendin-4 and GLP1 (7–36) protected HCAECs against lipoapoptosis, an effect that was blocked by PKA, PI3K/Akt, eNOS, p38 MAPK, and JNK inhibitors. Genetic silencing of eNOS also abolished the anti-apoptotic effect afforded by exendin-4. Our results support the notion that GLP1 receptor agonists restore eNOS-induced ROS production due to lipotoxicity and that such agonists protect against lipoapoptosis through PKA-PI3K/Akt-eNOS-p38 MAPK-JNK-dependent pathways via a GLP1 receptor-dependent mechanism.

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