High glucose-induced apoptosis in human coronary artery endothelial cells involves up-regulation of death receptors

IntroductionDiabetes-associated endothelium dysfunction might be linked to disturbances in Ca2+ homeostasis. Our main objective is to reveal the potential mechanisms by which high-glucose (HG) exposure promotes increased proliferation of human coronary artery endothelial cells (HCAECs) in culture, and that store-operated Ca2+ entry (SOCE) and insulin-like growth factor binding protein 3 (IGFBP3) contribute to this proliferation.Research design and methodsWe detected the expression levels of Ca2+ release-activated calcium channel proteins (Orais), IGFBP3 and proliferating cell nuclear antigen of HCAECs cultured in HG medium for 1, 3, 7, and 14 days and in streptozotocin-induced diabetic mouse coronary endothelial cells. Coimmunoprecipitation and immunofluorescence technologies were used to detect the interactions between Orais and IGFBP3 of HCAECs exposed to HG environment, and to detect IGFBP3 expression and proliferation after treatment of HCAECs cultured in HG medium with an agonist or inhibitor of SOCE. Similarly, after transfection of specific small interfering RNA to knock down IGFBP3 protein expression, SOCE activity and Orais expression were tested. Some processes related to endothelial dysfunction, such as migration, barrier function and adhesion marker expression, are also measured.ResultsHG exposure promoted increased proliferation of HCAECs in culture and that SOCE and IGFBP3 contributed to this proliferation. In addition, we also found that Orais and IGFBP3 were physically associated and regulated each other’s expression levels. Besides, their expression levels and interactions were enhanced in HCAECs after exposure to HG. HG exposure promotes cell migration, but reduces barrier function and adherens junction protein expression levels in HCAECs.ConclusionOrais and IGFBP3 formed a signaling complex that mediated HCAEC proliferation during HG exposure in culture. Meanwhile, we also found that SOCE stimulates proliferation of HCAECs by regulating IGFBP3, thereby promoting the occurrence and progression of coronary atherosclerosis in diabetes. It is worth noting that our findings may shed new light on the mechanisms of increased proliferation in HCAECs in diabetes and suggest the potential value of SOCE and IGFBP3 as therapeutic targets for coronary atherosclerosis in individuals with diabetes.

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GW25-e4422 S1P/S1PR2 mediated vascular endothelial cells dysfunction through upregulating miRNA-31 expression in human coronary artery endothelial cells exposed to high glucose

Journal of the American College of Cardiology ◽

10.1016/j.jacc.2014.06.069 ◽

2014 ◽

Vol 64 (16) ◽

pp. C13

Author(s):

Liu Weihua ◽

Liu Shiming

Keyword(s):

Endothelial Cells ◽

Coronary Artery ◽

High Glucose ◽

Vascular Endothelial Cells ◽

Vascular Endothelial ◽

Human Coronary Artery

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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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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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