Oxyhemoglobin Produces Necrosis in Cultured Smooth Muscle Cells

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Brain Edema XI

Part of the book series: Acta Neurochirurgica Supplements ((NEUROCHIRURGICA,volume 76))

Abstract

Object. Myonecrosis in the tunica media, which is defined morphologically, is one of the most striking alterations in the cerebral arterial wall following subarachnoid hemorrhage (SAH). In this study, oxyhemoglobin (OxyHb) was added to cultured rat aortic smooth muscle cells to determine the pattern of cell death by morphological and biochemical techniques.

Methods. Confluent rat aortic smooth muscle 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 culture dishes after exposed to OxyHb. To identify cell death pattern, DNA analysis, electron microscopy, and Western blotting using poly (ADP-ribose) polymerase (PARP) antibody were performed.

Conclusions. OxyHb decreased cell density in a concentration- and time-dependent manner. DNA analysis showed a smear pattern characteristic of cell necrosis. Transmission electron microscopy demonstrated disintegration of cell membrane and destruction of cell organelles. No apoptotic changes, such as condensation of chromatin or apoptotic bodies were observed. Western blotting using PARP antibody revealed that 116 kDa PARP was not cleaved to 85 kDa, an apoptosis-related fragment. These results demonstrated morphologically and biochemically that OxyHb induced necrosis, not apoptosis, in cultured smooth muscle cells.

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References

  1. Findlay JM, Weir BK, Kanamaru K, Espinosa F (1989) Arterial wall changes in cerebral vasospasm. Neurosurgery 25: 736–745

    Article  PubMed  CAS  Google Scholar 

  2. Fujii S, Fujitsu K (1988) Experimental vasospasm in cultured arterial smooth-muscle cells. Part 1: contractile and ultra-structural changes caused by oxyhemoglobin. J Neurosurg 69: 92–97

    Article  PubMed  CAS  Google Scholar 

  3. Li PF, Dietz R, von Harsdorf R (1997) Differential effect of hydrogen peroxide and superoxide anion on apoptosis and proliferation of vascular smooth muscle cells. Circulation 96: 3602–3609

    Article  PubMed  CAS  Google Scholar 

  4. Macdonald RL, Weir BK (1991) A review of hemoglobin and the pathogenesis of cerebral vasospasm. Stroke 22: 971–982

    Article  PubMed  CAS  Google Scholar 

  5. Martin W, Villani GM, Jothianandan D, Furchgott RF (1985) Selective blockade of endothelium-dependent and glyceryl trini-trate-induced relaxation by hemoglobin and by methylene blue in the rabbit aorta. J Pharm Exp Ther 232: 708–716

    CAS  Google Scholar 

  6. Mayberg MR, Okada T, Bark DH (1990) The role of hemoglobin in arterial narrowing after subarachnoid hemorrhage. J Neurosurg 72: 634–640

    Article  PubMed  CAS  Google Scholar 

  7. Oka Y, Ohta S, Todo H, Kohno K, Kumon Y, Sakaki S (1996) Protein synthesis and immunoreactivities of contraction-related proteins in smooth muscle cells of canine basilar artery after experimental subarachnoid hemorrhage. J Cereb Blood Flow Metab 16: 1335–1344

    Article  PubMed  CAS  Google Scholar 

  8. Okada H, Endo S, Kamiyama K, Suzuki J (1980) Oxyhemoglobin-induced cerebral vasospasm and sequential changes of vascular ultrastructure. Neurol Med Chir 20: 573–582

    Article  CAS  Google Scholar 

  9. Pluta RM, Afshar JK, Boock RJ, Oldfield EH (1998) Temporal changes in perivascular concentrations of oxyhemoglobin, deoxyhemoglobin, and methemoglobin after subarachnoid hemorrhage. J Neurosurg 88: 557–561

    Article  PubMed  CAS  Google Scholar 

  10. Steele JA, Stockbridge N, Maljkovic G, Weir B (1991) Free radicals mediate actions of oxyhemoglobin on cerebrovascular smooth muscle cells. Circ Res 68: 416–423

    Article  PubMed  CAS  Google Scholar 

  11. Takenaka K, Kassell NF, Foley PL, Lee KS (1993) Oxyhemoglobin-induced cytotoxicity and arachidonic acid release in cultured bovine endothelial cells. Stroke 24: 839–845

    Article  PubMed  CAS  Google Scholar 

  12. Tsukada T, Eguchi K, Migita K, Kawabe Y, Kawakami A, Matsuoka N, Takashima H, Mizokami A, Nagataki S (1995) Transforming growth factor beta 1 induces apoptotic cell death in cultured human umbilical vein endothelial cells with down-regulated expression of bcl-2. Biochem Biophys Res Comm 210: 1076–1082

    Article  PubMed  CAS  Google Scholar 

  13. Yoshimoto Y, Kim P, Sasaki T, Kirino T, Takakura K (1995) Functional changes in cultured strips of canine cerebral arteries after prolonged exposure to oxyhemoglobin. J Neurosurg 83: 867–874

    Article  PubMed  CAS  Google Scholar 

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© 2000 Springer-Verlag Wien

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Ogihara, K., Zubkov, A.Y., Parent, A.D., Zhang, J.H. (2000). Oxyhemoglobin Produces Necrosis in Cultured Smooth Muscle Cells. In: Mendelow, A.D., et al. Brain Edema XI. Acta Neurochirurgica Supplements, vol 76. Springer, Vienna. https://doi.org/10.1007/978-3-7091-6346-7_106

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  • DOI: https://doi.org/10.1007/978-3-7091-6346-7_106

  • Publisher Name: Springer, Vienna

  • Print ISBN: 978-3-7091-7257-5

  • Online ISBN: 978-3-7091-6346-7

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