Oxyhemoglobin produces apoptosis and necrosis in cultured smooth muscle cells

• Published in: Brain research 2001-01, Vol.889 (1), p.89-97
• Main Authors: Ogihara, Kotaro, Aoki, Kazuya, Zubkov, Alexander Y, Zhang, John H
• Format: Article
• Language: English
• Subjects: Animals; Aorta, Thoracic - drug effects; Aorta, Thoracic - pathology; Apoptosis; Apoptosis - drug effects; Blotting, Western; Cell Survival - drug effects; DNA - analysis; In Situ Nick-End Labeling; Microscopy, Electron; Muscle, Smooth, Vascular - drug effects; Muscle, Smooth, Vascular - pathology; Necrosis; Oxyhemoglobin; Oxyhemoglobins - metabolism; Poly(ADP-ribose) Polymerases - immunology; Rats; Smooth muscle; subarachnoid hemorrhage; Vasospasm
• ISSN: 0006-8993; 1872-6240
• DOI: 10.1016/S0006-8993(00)03120-6

Confluent rat aortic smooth muscle cells were treated with OxyHb in a concentration- and time-dependent manner. A high concentration of OxyHb (100 μM) within 24 h decreased cell density. DNA analysis showed a smear pattern characteristic of cell necrosis. Transmission electron microscopy demonstrated disintegration of the cell membrane and destruction of cell organelles. Western blotting using PARP antibody revealed that 116 kDa PARP was not cleaved to 85 kDa, an apoptosis-related fragment. On the contrary, a low concentration of OxyHb (10 μM) produced apoptotic cell death at 72 h that was supported by DNA analysis and TUNEL staining. These results demonstrated that a high level of OxyHb induced necrosis within 24 h and a low concentration of OxyHb produced apoptosis after 72 h in cultured smooth muscle cells. Morphological alterations induced by OxyHb might contribute to the vascular wall changes in the cerebral arteries following subarachnoid hemorrhage (SAH).