Adenoviral transfer of the heme oxygenase-1 gene protects striatal astrocytes from heme-mediated oxidative injury.
Heme oxygenase-1 (HO-1) is induced in the CNS after hemorrhage, and may have an effect on injury to surrounding tissue. Hemin, the preferred substrate of HO, is a neurotoxin that is present in intracranial hematomas. In a prior study, we observed that HO inhibitors increased the vulnerability of cultured cortical astrocytes to heme-mediated oxidative injury. To investigate the effect of HO more specifically, we used an adenoviral vector encoding the human HO-1 gene to specifically increase HO-1 expression. Incubation with 100 MOI of the HO-1 adenovirus (Adv-HHO-1) for 24 h increased both HO-1 protein and HO activity; a control adenovirus lacking the HO-1 gene had no effect. Using a DNA probe that was specific for human HO-1, 80.5 +/- 7.2% of astrocytes were observed to be infected by in situ hybridization. The cell death produced by 30-60 microM hemin was significantly reduced by pretreatment with 100 MOI Adv-HHO-1, as assessed by LDH release, propidium iodide exclusion, and MTT reduction assay. The threefold increase in cell protein oxidation produced by hemin was also attenuated in cultures pretreated with Adv-HHO-1. These results support the hypothesis that HO-1 protects astrocytes from heme-mediated oxidative injury. Specifically increasing astrocytic HO-1 by gene transfer may have a beneficial effect on hemorrhagic CNS injury.
Department of Emergency Medicine, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Zhi-Ping
ZP
Teng
Jing
J
Chen
Lee-Young
LY
Chau
Nicholas
N
Galunic
Raymond F
RF
Regan
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D002478:Cells, Cultured; D003342:Corpus Striatum / Q000187:drug effects / Q000473:pathology*; D004351:Drug Resistance; D005293:Ferritins / Q000378:metabolism; D018014:Gene Transfer Techniques; D005978:Glutathione / Q000378:metabolism; D006419:Heme Oxygenase (Decyclizing) / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D051547:Heme Oxygenase-1; D006427:Hemin / Q000494:pharmacology*; D006801:Humans; D008565:Membrane Proteins; D051379:Mice; D008815:Mice, Inbred Strains; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress; D016133:Polymerase Chain Reaction
D008565:Membrane Proteins; D006427:Hemin; D005293:Ferritins; C496541:HMOX1 protein, human; D006419:Heme Oxygenase (Decyclizing); D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; D005978:Glutathione