INABIS '98 - Oxyhemoglobin Induced Apoptosis In Cultured Endothelial Cell

Oxyhemoglobin Induced Apoptosis In Cultured Endothelial Cell
Zhang, J. (Department of Neurosurgery, University of Mississippi Medical Centre, USA)
Ogihara, K. (Department of Neurosurgery, University of Mississippi Medical Centre, USA)
Zubkov, AY. (Department of Neurosurgery, University of Mississippi Medical Center, USA)
Bernanke, DH. (Department of Anatomy, University of Mississippi Medical Center, USA)
Lewis, AI. (Department of Neurosurgery, University of Mississippi Medical Center, USA)
Parent, AD. (Department of Neurosurgery, University of Mississippi Medical Center, USA)

Contact Person: John Zhang, MD, PhD (jzhang@neurosurgery.umsmed.edu)

Abstract

Object: Oxyhemoglobin (OxyHb) is one of the most important spasmogens for cerebral vasospasm that follows aneurysmal subarachnoid hemorrhage (SAH). The cytotoxicity effect of OxyHb has been documented in endothelial and smooth muscle cells, however, the pattern of cell death, necrosis or apoptosis, as a 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 the cultre dishes after exposed to OxyHb. To identify apoptotic changes, three specific methods: DNA fragmentation (electrophoreses), Apoptotic bod (transmission electron microscopy) and cleavage of poly (ADP-ribose) polymerase (PARP) (Western Blotting) were employed. Conclusions: OxyHb decreased cell density in a concentration and time-dependent manner. DNA analysis 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 using PARP antibody revealed that 116 kDa PARP was cleaved to 85 kDa, an apoptosis-related fragment. These results for the first time demonstrated that OxyHb induced apoptosis in cultured endothelial cell.

Back to the top.
Presentation Number SAjohn zhang0443
Keywords: oxyhemoglobin, apoptosis, endothelial cell, vasospasm, western blotting

| Discussion Board | Next Page | Your Symposium |

Zhang, J.; Ogihara, K.; Zubkov, AY.; Bernanke, DH.; Lewis, AI.; Parent, AD.; (1998). Oxyhemoglobin Induced Apoptosis In Cultured Endothelial Cell. Presented at INABIS '98 - 5th Internet World Congress on Biomedical Sciences at McMaster University, Canada, Dec 7-16th. Invited Symposium. Available at URL http://www.mcmaster.ca/inabis98/zhang/zhang0443/index.html