Oxyhemoglobin-induced apoptosis in cultured endothelial cells
| Autor: | David H. Bernanke, Adam I. Lewis, John H. Zhang, Andrew D. Parent, Alexander Y. Zubkov, Kotaro Ogihara |
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| Rok vydání: | 1999 |
| Předmět: |
Time Factors
Endothelium Poly ADP ribose polymerase Blotting Western Apoptosis Cell Count DNA Fragmentation Biology Muscle Smooth Vascular Necrosis medicine Animals Aorta Cells Cultured Cell Nucleus Electrophoresis Agar Gel Cell Death Dose-Response Relationship Drug Intracranial Aneurysm Subarachnoid Hemorrhage Apoptotic body Electrophoreses Molecular biology Clone Cells Nucleosomes Endothelial stem cell Microscopy Electron medicine.anatomical_structure Ischemic Attack Transient Cell culture Oxyhemoglobins Luminescent Measurements Immunology DNA fragmentation Cattle Electrophoresis Polyacrylamide Gel Endothelium Vascular Poly(ADP-ribose) Polymerases |
| Zdroj: | Journal of Neurosurgery. 91:459-465 |
| ISSN: | 0022-3085 |
| DOI: | 10.3171/jns.1999.91.3.0459 |
| Popis: | Object. Oxyhemoglobin (OxyHb) is one of the most important spasmogens for cerebral vasospasm that follows aneurysmal subarachnoid hemorrhage. The cytotoxic effect of OxyHb has been documented in endothelial and smooth-muscle cells; however, the pattern of cell death—necrosis or apoptosis—as the final stage of cell damage has not been demonstrated. This study was undertaken to determine if OxyHb induces apoptotic changes in cultured bovine aortic endothelial cells.Methods. Confluent bovine aortic endothelial cells were treated with OxyHb in a concentration- and time-dependent manner. Cell density was assayed by counting the number of cells that attached to culture dishes after exposure to OxyHb. To identify apoptotic changes, the investigators used three specific methods: DNA fragmentation (electrophoreses), the apoptotic body (transmission electron microscopy), and cleavage of poly (adenosine diphosphate ribose) polymerase (PARP [Western blotting]).Conclusions. Oxyhemoglobin decreased cell density in a concentration- and time-dependent manner. Analysis of DNA showed a pattern of internucleosomal cleavage characteristic of apoptosis (DNA ladder). Transmission electron microscopy demonstrated condensation of nuclei and apoptotic bodies in OxyHb-treated endothelial cells. Western blotting with the PARP antibody revealed that the 116-kD PARP was cleaved to the 85-kD apoptosis-related fragment. These results for the first time demonstrated that the OxyHb induces apoptosis in cultured endothelial cells. |
| Databáze: | OpenAIRE |
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